supporting - royal society of chemistry · s1 supporting information exploration of the active site...

52
S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn, a# Karin Thorsheim, a# Jonas Ståhle, b Sophie Manner, a Christoffer Hamark, b Andrea Persson, c Emil Tykesson, c Katrin Mani, c Gunilla Westergren-Thorsson, c Göran Widmalm, b Ulf Ellervik a * a Center for Analysis and Synthesis, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, SE-221 00 Lund, Sweden b Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, SE-106 91 Stockholm, Sweden c Department of Experimental Medical Science, Lund University, BMC, SE-221 00 Lund, Sweden # These authors contributed equally to this work Table of Contents Mass analysis ............................................................................................................................ 3 Cloning, expression and purification of β 4GALT7 .............................................................. 4 1 H-NMR Phenyl β-D-xylopyranoside (3) ................................................................................. 5 13 C-NMR Phenyl β-D-xylopyranoside (3) ................................................................................ 6 1 H-NMR 1-Naphthyl β-D-xylopyranoside (4) .......................................................................... 7 13 C-NMR 1-Naphthyl β-D-xylopyranoside (4) ......................................................................... 8 1 H-NMR 4-Phenylphenyl β-D-xylopyranoside (5) ................................................................... 9 13 C-NMR 4-Phenylphenyl β-D-xylopyranoside (5) ................................................................ 10 1 H-NMR (9-Anthracene)-methyl β-D-xylopyranoside (6) ...................................................... 11 13 C-NMR (9-Anthracene)-methyl β-D-xylopyranoside (6)..................................................... 12 1 H-NMR 2-Naphthyl 1-thio-β-D-xylopyranoside (7) ............................................................. 13 13 C-NMR 2-Naphthyl 1-thio-β-D-xylopyranoside (7) ............................................................ 14 1 H-NMR 2-Naphthyl β-D-xylopyranosyl sulfone (8) ............................................................. 15 13 C-NMR 2-Naphthyl β-D-xylopyranosyl sulfone (8) ............................................................ 16 1 H-NMR 2-Naphthyl β-L-xylopyranoside (10) ....................................................................... 17 13 C-NMR 2-Naphthyl β-L-xylopyranoside (10) ...................................................................... 18 1 H-NMR 2-Naphthyl α-D-xylopyranoside (11) ...................................................................... 19 Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry. This journal is © The Royal Society of Chemistry 2015

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Page 1: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S1

Supporting Information

Exploration of the active site of β4GalT7: Modifications of the

aglycon of aromatic xylosides Anna Siegbahn,a# Karin Thorsheim,a# Jonas Ståhle,b Sophie Manner,a Christoffer Hamark,b

Andrea Persson,c Emil Tykesson,c Katrin Mani,c Gunilla Westergren-Thorsson,c Göran

Widmalm,b Ulf Ellervika*

a Center for Analysis and Synthesis, Center for Chemistry and Chemical Engineering, Lund University, P.O.

Box 124, SE-221 00 Lund, Sweden b Department of Organic Chemistry, Arrhenius Laboratory, Stockholm University, SE-106 91 Stockholm,

Sweden c Department of Experimental Medical Science, Lund University, BMC, SE-221 00 Lund, Sweden # These authors contributed equally to this work

Table of Contents Mass analysis ............................................................................................................................ 3

Cloning, expression and purification of β4GALT7 .............................................................. 4 1H-NMR Phenyl β-D-xylopyranoside (3) ................................................................................. 5 13C-NMR Phenyl β-D-xylopyranoside (3) ................................................................................ 6 1H-NMR 1-Naphthyl β-D-xylopyranoside (4) .......................................................................... 7 13C-NMR 1-Naphthyl β-D-xylopyranoside (4) ......................................................................... 8 1H-NMR 4-Phenylphenyl β-D-xylopyranoside (5) ................................................................... 9 13C-NMR 4-Phenylphenyl β-D-xylopyranoside (5) ................................................................ 10 1H-NMR (9-Anthracene)-methyl β-D-xylopyranoside (6) ...................................................... 11 13C-NMR (9-Anthracene)-methyl β-D-xylopyranoside (6) ..................................................... 12 1H-NMR 2-Naphthyl 1-thio-β-D-xylopyranoside (7) ............................................................. 13 13C-NMR 2-Naphthyl 1-thio-β-D-xylopyranoside (7) ............................................................ 14 1H-NMR 2-Naphthyl β-D-xylopyranosyl sulfone (8) ............................................................. 15 13C-NMR 2-Naphthyl β-D-xylopyranosyl sulfone (8) ............................................................ 16 1H-NMR 2-Naphthyl β-L-xylopyranoside (10) ....................................................................... 17 13C-NMR 2-Naphthyl β-L-xylopyranoside (10) ...................................................................... 18 1H-NMR 2-Naphthyl α-D-xylopyranoside (11) ...................................................................... 19

Electronic Supplementary Material (ESI) for Organic & Biomolecular Chemistry.This journal is © The Royal Society of Chemistry 2015

Page 2: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S2

13C-NMR 2-Naphthyl α-D-xylopyranoside (11) ..................................................................... 20 1H-NMR 2-(6-Cyano-naphthyl) β-D-xylopyranoside (19) ..................................................... 21 13C-NMR 2-(6-Cyano-naphthyl) β-D-xylopyranoside (19) .................................................... 22 1H-NMR 2-Naphthyl 1-thio-α-L-arabinopyranoside (23) ....................................................... 23 13C-NMR 2-Naphthyl 1-thio-α-L-arabinopyranoside (23) ...................................................... 24 1H-NMR α-L-Arabinopyranoside 2-naphthyl sulfone (24) ..................................................... 25 13C-NMR α-L-Arabinopyranoside 2-naphthyl sulfone (24) ................................................... 26 1H-NMR (2-Naphthyl)-methyl α-L-arabinopyranoside (25) .................................................. 27 13C-NMR (2-Naphthyl)-methyl α-L-arabinopyranoside (25) ................................................. 28 1H-NMR 5-(2-Naphthoxy)-3-oxo-1-pentyl α-L-arabinopyranoside (26) ............................... 29 13C-NMR 5-(2-Naphthoxy)-3-oxo-1-pentyl α-L-arabinopyranoside (26) .............................. 30 1H-NMR 1-Naphthyl 2,3,4-tri-O-acetyl-β-D-xylopyranoside (29) ......................................... 31 13C-NMR 1-Naphthyl 2,3,4-tri-O-acetyl-β-D-xylopyranoside (29) ........................................ 32 1H-NMR 4-Phenylphenyl 2,3,4-tri-O-acetyl-β-D-xylopyranoside (30) .................................. 33 13C-NMR 4-Phenylphenyl 2,3,4-tri-O-acetyl-β-D-xylopyranoside (30) ................................. 34 1H-NMR 2-(6-Cyano-naphthyl) 2,3,4-tri-O-acetyl-β-D-xylopyranoside (31) ........................ 35 13C-NMR 2-(6-Cyano-naphthyl) 2,3,4-tri-O-acetyl-β-D-xylopyranoside (31) ....................... 36 1H-NMR 2-Naphthyl 2,3,4-tri-O-acetyl-1-thio-β-D-xylopyranoside (32) .............................. 37 13C-NMR 2-Naphthyl 2,3,4-tri-O-acetyl-1-thio-β-D-xylopyranoside (32) ............................. 38 1H-NMR 2-Naphthyl 2,3,4-tri-O-acetyl β-D-xylopyranosyl sulfone (33) .............................. 39 13C-NMR 2-Naphthyl 2,3,4-tri-O-acetyl β-D-xylopyranosyl sulfone (33) ............................. 40 1H-NMR 2-Naphthyl 2,3,4-tri-O-acetyl β-L-xylopyranoside (35) .......................................... 41 13C-NMR 2-Naphthyl 2,3,4-tri-O-acetyl β-L-xylopyranoside (35) ......................................... 42 1H-NMR 2-Naphthyl 2,3,4-tri-O-acetyl 1-thio-α-L-arabinopyranoside (38) .......................... 43 13C-NMR 2-Naphthyl 2,3,4-tri-O-acetyl 1-thio-α-L-arabinopyranoside (38) ......................... 44 1H-NMR 2-Naphthyl 2,3,4-tri-O-acetyl α-L-arabinopyranosyl sulfone (39) .......................... 45 13C-NMR 2-Naphthyl 2,3,4-tri-O-acetyl α-L-arabinopyranosyl sulfone (39) ........................ 46 1H-NMR (2-Naphthyl)-methyl 2,3,4-tri-O-acetyl α-L-arabinopyranoside (40) ..................... 47 13C-NMR (2-Naphthyl)-methyl 2,3,4-tri-O-acetyl α-L-arabinopyranoside (40) .................... 48 1H-NMR 5-Chloro-3-oxo-1-pentyl 2,3,4-tri-O-acetyl-α-L-arabinopyranoside (42) ............... 49 13C-NMR 5-Chloro-3-oxo-1-pentyl 2,3,4-tri-O-acetyl-α-L-arabinopyranoside (42) .............. 50 1H-NMR 5-(2-Naphthoxy)-3-oxo-1-pentyl 2,3,4-tri-O-acetyl-α-L-arabinopyranoside (43) .. 51 13C-NMR 5-(2-Naphthoxy)-3-oxo-1-pentyl 2,3,4-tri-O-acetyl-α-L-arabinopyranoside (43) . 52

 

Page 3: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S3

Mass analysis Table S1. HRMS for all galactosylated products.

a HRMS for [M+NH4]+

Acceptor substrate

Galactosylation product Chemical formula

HRMS calcd for [M+H]+

HRMS found

1 C21H26O10 439.1599 439.1598 2 C12H22O10 327.1286 327.1287 3 C17H24O10 389.1442 389.1441 4 C21H26O10 439.1599 439.1598 5 C23H28O10 465.1755 465.1755 6 C26H30O10 520.2177a 520.2179 7 C21H26O9S 455.1370 455.1370 8 C21H26O11S 487.1269 487.1270 9 C22H28O9 437.1806 437.1804 12 C22H28O10 453.1755 453.1755 13 C23H30O11 483.1861 483.1858 14 C25H34O12 527.2123 527.2123 15 C27H38O13 571.2385 571.2388 16 C22H28O11 486.1973a 486.1972 17 C21H26O11 455.1548 455.1549 18 C23H28O12 497.1654 497.1655 19 C22H25NO10 464.1551 464.1550

Page 4: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S4

Cloning, expression and purification of β4GALT7 The part of the b4GALT7 open reading frame (sequence harmonized for expression in E.

Coli, Genewiz) corresponding to aa 82 to aa 327 was subcloned into the NcoI and NotI sites

of a pET-41a(+) (Merck Millipore) expression vector using the following primers (Sigma):

Forward (NcoI restriction site in bold letters)

aa 82: 5’-GCATCTCCCATGGAGCATTGGGAGGAAGATGCCAGTT-3'

Reverse (NotI restriction site in bold letters) aa 327: 5’-

GCATCTGCGGCCGCATTAGCTAAATGTACACCAAGGGGTTGC-3’

The plasmid product, pET-41a(+)-β4GALT7aa82, was amplified (Library Efficiency DH5α

bacteria, Invitrogen), purified (HiSpeed Plasmid Midi Kit, Qiagen) and sequenced (Eurofins

MWG Operon). BL21(DE3) E. Coli bacteria (Merck Millipore) were transformed with pET-

41a(+)-β4GALT7aa82 and grown on LB agar containing kanamycin (30 mg/ml). A resistant

colony was transferred to selective LB medium and grown at 37 °C with shaking at 250 rpm

to OD600≈0.5. Expression was induced at 32 °C with IPTG (0.56 mM, Sigma) and culture

was left shaking at 250 rpm over night. Cells were pelleted and then lysed with Cellytic (1X,

Sigma) in Tris (20 mM, pH 7.6), NaCl (500 mM), Benzonase (50 U/ml, Sigma), lysozyme

(0.2 mg/ml, Sigma) and PMSF (1 mM) for 20 min at 22 °C. Clarified lysate was applied to an

equilibrated HisTrap HP column (GE Healthcare) at a flowrate of 1 ml/min. The column was

washed with a phosphate buffer (20 mM, pH 7.4) containing NaCl (500 mM) and imidazole

(40 mM), after which the protein was eluted by an imidazole gradient (40-250 mM). Protein

purity for each eluted fraction was analyzed by SDS-PAGE on a Bis-Tris 4-12 % gel in

MOPS buffer (Invitrogen), visualized by Brilliant Blue G colloidal (Sigma). Fractions

containing pure protein were pooled and buffer exchanged into TBS (20 mM, pH 7.9) on PD-

10 columns (GE Healthcare).  

Page 5: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S5

1H-NMR Phenyl β-D-xylopyranoside (3)

 

23

45

67

89

10

ppm3.353

3.3793.4023.4233.4303.4383.4443.4473.4503.4703.5483.5563.5613.5703.5733.5763.5833.5873.5953.6023.6093.8943.9073.9223.9364.8504.8564.8624.8696.9826.9846.9877.0017.0037.0057.0197.0217.0247.0307.0377.0397.0427.0447.0537.0587.0617.0647.0667.2497.2557.2607.2687.2737.2777.2837.2907.2937.2957.301

1.081.980.99

1.00

1.09

1.001.941.96

TD0 1

D1 0.00100000 sec

TE 293.4 K

DE 6.50 usec

DW 69.333 usec

RG 71.8

AQ 3.9998901 sec

FIDRES 0.125005 Hz

SWH 7211.539 Hz

DS 2

NS 32

SOLVENT MeOD

TD 57690

PULPROG zg30

PROBHD 5 mm PABBO BB−

INSTRUM spect

Time 15.30

Date_ 20141209

PROCNO 1

EXPNO 1

NAME KT−D150

OHO H

OOHO

3

Page 6: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S6

13C-NMR Phenyl β-D-xylopyranoside (3)

2030

4050

6070

8090

100

110

120

130

140

150

160

170

180

190

200

210

ppm

66.85970.99074.71177.679

102.815

117.750

123.445

130.393

158.894

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

6322

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

93.8

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

8

000

SOLV

ENT

M

eOD

TD

480

74PU

LPRO

G

zgd

c30

PRO

BHD

5

mm

PA

BBO

BB−

INST

RUM

s

pect

Tim

e

2

2.27

Dat

e_

201

4120

9PR

OCN

O

1EX

PNO

3

NA

ME

K

T−D

150

OHO H

OOHO

3

Page 7: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S7

1H-NMR 1-Naphthyl β-D-xylopyranoside (4)

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

ppm

3.3783.4043.4063.4323.4593.4813.5033.6043.6173.6293.6363.6393.6513.6583.6643.9423.9553.9703.983

5.0545.072

7.1287.1477.3627.3827.4027.4427.4557.4607.4707.4797.4857.4977.5027.5147.5357.7997.8087.8167.8228.3488.3558.3628.372

1.1571.0882.098

1.063

1.074

1.049

1.1263.135

1.027

1.000

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0078

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.3 K

DE

6

.50

usec

DW

69

.333

use

cRG

203

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

32

SOLV

ENT

M

eOD

TD

576

90PU

LPRO

G

zg

30PR

OB

HD

5

mm

PA

BB

O B

B−

INST

RUM

s

pect

Tim

e

1

3.47

Dat

e_

201

4101

4PR

OCN

O

1EX

PNO

2

NA

ME

SM

−D11

5_D

OHO H

OOHO

4

Page 8: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S8

13C-NMR 1-Naphthyl β-D-xylopyranoside (4)

3040

5060

7080

90100

110

120

130

140

150

160

170

180

190

200

210

ppm

66.980

71.07174.85777.828

103.276

110.451

123.149123.291126.335126.806127.349127.456128.439136.056

154.471

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

6270

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.4

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

4

000

SOLV

ENT

M

eOD

TD

480

74PU

LPRO

G

zgd

c30

PRO

BHD

5

mm

PA

BBO

BB−

INST

RUM

s

pect

Tim

e

0

.40

Dat

e_

201

4101

5PR

OCN

O

1EX

PNO

3

NA

ME

SM

−D11

5_D

OHO H

OOHO

4

Page 9: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S9

1H-NMR 4-Phenylphenyl β-D-xylopyranoside (5)

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

ppm

3.3563.3823.3853.4103.4323.4443.4493.4533.4623.4843.5773.5813.5913.5943.6163.9153.9283.9433.957

4.8984.916

7.1137.1207.1257.1377.1427.1497.2667.2697.2877.3037.3057.3817.4017.4157.4197.5287.5357.5407.5537.5577.5657.5737.576

3.1961.049

1.043

1.369

2.0731.0002.0794.168

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0079

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.2 K

DE

6

.50

usec

DW

69

.333

use

cRG

181

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

32

SOLV

ENT

M

eOD

TD

576

90PU

LPRO

G

zg

30PR

OB

HD

5

mm

PA

BB

O B

B−

INST

RUM

s

pect

Tim

e

8

.57

Dat

e_

201

4091

1PR

OCN

O

1EX

PNO

4

NA

ME

SM

−D12

3_D

OHO H

OOHO

5

Page 10: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S10

13C-NMR 4-Phenylphenyl β-D-xylopyranoside (5)

3040

5060

7080

90100

110

120

130

140

150

160

170

180

190

200

210

ppm

66.960

71.04974.77477.749

102.889

118.098

127.673127.893128.952129.798

136.792

141.965

158.473

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

6272

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.6

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

1

500

SOLV

ENT

M

eOD

TD

480

74PU

LPRO

G

zgd

c30

PRO

BHD

5

mm

PA

BBO

BB−

INST

RUM

s

pect

Tim

e

7

.32

Dat

e_

201

4091

2PR

OCN

O

1EX

PNO

6

NA

ME

SM

−D12

3_D

OHO H

OOHO

5

Page 11: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S11

1H-NMR (9-Anthracene)-methyl β-D-xylopyranoside (6)

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

ppm3.181

3.2003.2043.2183.2233.2443.2463.2503.2723.2943.4853.5143.5273.5353.5393.5493.5533.5613.5743.5953.6493.9914.0054.0204.0334.4424.461

5.6515.6795.8195.847

7.4647.4667.4807.4827.4877.5017.5037.5367.5407.5527.5587.5627.5757.5788.0358.0568.4808.5028.530

3.354

1.274

0.988

0.941

1.0161.000

1.9602.146

2.015

2.985

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0078

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

295

.1 K

DE

6

.50

usec

DW

69

.333

use

cRG

181

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

128

SOLV

ENT

M

eOD

TD

576

90PU

LPRO

G

zg

30PR

OB

HD

5

mm

PA

BB

O B

B−

INST

RUM

s

pect

Tim

e

1

6.51

Dat

e_

201

4091

5PR

OCN

O

1EX

PNO

1

NA

ME

SM

−D13

7_C

OHO H

OOHO

6

Page 12: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S12

13C-NMR (9-Anthracene)-methyl β-D-xylopyranoside (6)

3040

5060

7080

90100

110

120

130

140

150

160

170

180

190

200

210

ppm

63.77067.184

71.29874.83377.888

103.997

125.607126.099127.323128.985129.635129.948132.588132.904

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

6271

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.9

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

5

000

SOLV

ENT

M

eOD

TD

480

74PU

LPRO

G

zgd

c30

PRO

BHD

5

mm

PA

BBO

BB−

INST

RUM

s

pect

Tim

e

2

.30

Dat

e_

201

4091

8PR

OCN

O

1EX

PNO

3

NA

ME

SM

−D13

7_C

OHO H

OOHO

6

Page 13: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S13

1H-NMR 2-Naphthyl 1-thio-β-D-xylopyranoside (7)

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

ppm

3.2293.2493.2513.2743.3503.3723.3933.4553.4683.4783.4813.4903.4933.5023.5153.9543.9663.9823.9944.6564.679

7.4517.4647.4697.4797.4887.4947.5067.5817.5857.6037.6077.7907.8127.8197.8267.8337.8428.0098.011

1.8201.3371.069

0.999

1.000

1.9531.024

3.062

1.039

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0080

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.2 K

DE

6

.50

usec

DW

69

.333

use

cRG

161

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

8

SOLV

ENT

M

eOD

TD

576

90PU

LPRO

G

zg

30PR

OB

HD

5

mm

PA

BB

O B

B−

INST

RUM

s

pect

Tim

e

1

1.13

Dat

e_

201

4082

7PR

OCN

O

1EX

PNO

1

NA

ME

SM

−D12

1_D

OHO H

OOHS

7

Page 14: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S14

13C-NMR 2-Naphthyl 1-thio-β-D-xylopyranoside (7)

3040

5060

7080

90100

110

120

130

140

150

160

170

180

190

200

210

ppm

70.46670.90973.758

79.221

90.010

127.321127.598128.500128.690129.332130.514131.848132.234133.956135.050

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

6280

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.6

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

5

12SO

LVEN

T

MeO

DTD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B−IN

STRU

M

spe

ctTi

me

11.

34D

ate_

2

0140

827

PRO

CNO

1

EXPN

O

2N

AM

E

SM−D

121_

D

OHO H

OOHS

7

Page 15: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S15

1H-NMR 2-Naphthyl β-D-xylopyranosyl sulfone (8)

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

10.0

10.5

ppm

3.1453.1603.1713.1813.1913.1993.3473.3633.3693.3803.3873.3943.4853.6253.6473.6703.8503.8623.8783.8904.4304.453

7.6557.6587.6757.6937.6957.7117.7147.7327.7497.7527.8847.8887.9067.9108.0128.0338.0698.0908.1088.540

1.2042.1310.2010.987

1.072

0.046

1.0000.051

2.0121.0141.0062.036

0.984PC

4

.00

GB

0LB

0

.30

Hz

SSB

0W

DW

EMSF

4

00.1

3000

78 M

Hz

SI

1310

72P1

9

.80

usec

NU

C1

1H

SFO

1

4

00.1

3320

10 M

Hz

====

====

CH

AN

NEL

f1 =

====

===

TD0

1D

1

0.0

0100

000

sec

TE

294

.7 K

DE

6

.50

usec

DW

69

.333

use

cRG

203

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

32

SOLV

ENT

M

eOD

TD

576

90PU

LPRO

G

zg

30PR

OB

HD

5

mm

PA

BB

O B

B−

INST

RUM

s

pect

Tim

e

1

2.27

Dat

e_

201

4091

2PR

OCN

O

1EX

PNO

1

NA

ME

K

T−D

090

OHO H

OOHS

OO

8

Page 16: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S16

13C-NMR 2-Naphthyl β-D-xylopyranosyl sulfone (8)

3040

5060

7080

90100

110

120

130

140

150

160

170

180

190

200

210

ppm

70.22471.03371.237

79.099

93.748

125.208128.754129.052130.035130.591130.633132.602133.492135.276137.047

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

6271

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.8

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

9

000

SOLV

ENT

M

eOD

TD

480

74PU

LPRO

G

zgd

c30

PRO

BHD

5

mm

PA

BBO

BB−

INST

RUM

s

pect

Tim

e

1

.27

Dat

e_

201

4091

4PR

OCN

O

1EX

PNO

3

NA

ME

K

T−D

090

OHO H

OOHS

OO

8

Page 17: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S17

1H-NMR 2-Naphthyl β-L-xylopyranoside (10)

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

ppm

3.4263.4413.4523.4553.4643.4803.4853.4873.5033.5073.5263.5793.5923.6003.6043.6143.6173.6263.6393.9543.9673.9823.9955.0195.037

7.2527.2587.2747.2807.3307.3337.3477.3507.3537.3677.3717.4117.4187.4287.4317.4357.4497.4527.7557.7757.798

3.1411.063

1.012

1.000

0.9761.0292.140

3.160

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0078

MH

zSI

13

1072

SFO

1

4

00.1

3340

11 M

Hz

PL1W

27.

0071

6019

WPL

1

5.00

dB

P1

9.1

0 us

ecN

UC1

1

H==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

300

.0 K

DE

6

.00

usec

DW

69

.333

use

cRG

362

AQ

3.99

9890

1 se

cFI

DR

ES

0.12

5005

Hz

SWH

7211

.539

Hz

DS

2N

S

32

SOLV

ENT

M

eOD

TD

576

90PU

LPRO

G

zg

30PR

OB

HD

5

mm

PA

BB

O B

B/

INST

RUM

s

pect

Tim

e

8

.44

Dat

e_

201

3081

2PR

OCN

O

1EX

PNO

1

NA

ME

K

H−C

002

OO

HOHO

OH

10

Page 18: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S18

13C-NMR 2-Naphthyl β-L-xylopyranoside (10)

2030

4050

6070

8090

100

110

120

130

140

150

160

170

180

190

200

210

ppm

67.00371.07474.81677.751

102.898

111.867

119.994125.267127.398128.125128.614130.356131.303135.833

156.662

SF

100

.612

6271

MH

zSI

3

2768

SFO

2

4

00.1

3160

05 M

Hz

PL12

W

0

.349

5255

4 W

PL2W

27.

0071

6019

WPL

12

1

3.88

dB

PL2

−5.

00 d

BPC

PD2

80.0

0 us

ecN

UC2

1

HCP

DPR

G2

wal

tz16

====

====

CH

AN

NEL

f2 =

====

===

SFO

1

1

00.6

2282

98 M

Hz

PL1W

73.

8220

0623

WPL

1

4.00

dB

P1

8.0

0 us

ecN

UC1

1

3C==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

3

00.0

KD

E

6.0

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

9

000

SOLV

ENT

M

eOD

TD

480

74PU

LPRO

G

zgd

c30

PRO

BHD

5

mm

PA

BBO

BB/

INST

RUM

s

pect

Tim

e

2

2.50

Dat

e_

201

3081

2PR

OCN

O

1EX

PNO

3

NA

ME

K

H−C

002

OO

HOHO

OH

10

Page 19: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S19

1H-NMR 2-Naphthyl α-D-xylopyranoside (11)

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

10.0

10.5

ppm

3.5943.6023.6053.6113.6253.6343.6633.6743.8373.8573.880

5.6155.624

7.2957.3017.3177.3247.3457.3637.3657.4047.4077.4247.4427.4447.4927.4987.7317.7527.7787.7937.801

4.05

0.96

1.00

2.021.041.00

3.04

PC 4.00

GB 0

LB 0.30 Hz

SSB 0

WDW EM

SF 400.1300078 MHz

SI 131072

P1 9.80 usec

NUC1 1H

SFO1 400.1332010 MHz

======== CHANNEL f1 ========

TD0 1

D1 0.00100000 sec

TE 293.6 K

DE 6.50 usec

DW 69.333 usec

RG 128

AQ 3.9998901 sec

FIDRES 0.125005 Hz

SWH 7211.539 Hz

DS 2

NS 8

SOLVENT MeOD

TD 57690

PULPROG zg30

PROB

HD

5 m

m PA

BBO

BB−

INSTRUM spect

Time 8.10

Date_ 20141110

PROCNO 1

EXPNO 1

NAME

KT

−alf

aXyl

Nap

OHO H

OHO

O

11

Page 20: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S20

13C-NMR 2-Naphthyl α-D-xylopyranoside (11)

2030

4050

6070

8090

100

110

120

130

140

150

160

170

180

190

200

210

ppm

63.713

71.39873.40375.160

99.141

111.924

120.106125.222127.381128.087128.612130.399131.198135.860

156.107

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

6285

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

93.9

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

5

12SO

LVEN

T

MeO

DTD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B−IN

STRU

M

spe

ctTi

me

8.2

9D

ate_

2

0141

110

PRO

CNO

1

EXPN

O

3N

AM

E

KT−

alfa

Xyl

Nap

OHO H

OHO

O

11

Page 21: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S21

1H-NMR 2-(6-Cyano-naphthyl) β-D-xylopyranoside (19)

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

ppm

3.3023.3063.3103.3143.3183.3493.4583.4663.4813.4913.4943.5013.5053.5073.5193.5243.5463.5883.6003.6223.9613.9743.9894.002

5.0955.113

7.3797.3857.4027.4087.4937.4987.5867.5907.6077.6117.9067.9247.9298.278

3.1211.043

1.033

1.019

0.017

0.018

1.0271.0321.017

2.065

1.000

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0079

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.7 K

DE

6

.50

usec

DW

69

.333

use

cRG

128

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

8

SOLV

ENT

M

eOD

TD

576

90PU

LPRO

G

zg

3035

2%+'�����PP�3$%%2�%%ï

INST

RUM

s

pect

Tim

e

1

4.13

Dat

e_

201

4090

5PR

OCN

O

1EX

PNO

1

NA

ME

S

MD

127B

OHO H

OOHO

CN19

Page 22: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S22

13C-NMR 2-(6-Cyano-naphthyl) β-D-xylopyranoside (19)

3040

5060

7080

90100

110

120

130

140

150

160

170

180

190

200

210

ppm

67.03870.96374.66377.638

102.351

108.317111.571

120.303121.847127.882129.560129.917131.273135.007137.618

159.089

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

6287

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

95.2

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

5

12SO

LVEN

T

MeO

DTD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B−IN

STRU

M

spe

ctTi

me

14.

42D

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0140

905

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CNO

1

EXPN

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2N

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7B

OHO H

OOHO

CN19

Page 23: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S23

1H-NMR 2-Naphthyl 1-thio-α-L-arabinopyranoside (23)

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

ppm

3.6003.6083.6163.6203.6283.6473.6513.7293.7503.7703.8963.9003.9043.9093.9133.9183.9994.0084.0304.0394.7564.777

7.4327.4367.4497.4537.4597.4667.4727.4797.4837.4957.5007.5837.5877.6047.6097.7797.8017.8097.8147.8328.0058.008

2.1691.0391.0311.077

1.000

2.1301.120

3.244

1.102

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0077

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.8 K

DE

6

.50

usec

DW

69

.333

use

cRG

181

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

32

SOLV

ENT

M

eOD

TD

576

90PU

LPRO

G

zg

3035

2%+'�����PP�3$%%2�%%ï

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RUM

s

pect

Tim

e

1

2.13

Dat

e_

201

4091

2PR

OCN

O

1EX

PNO

1

1$0(������������.

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OHS

OH

23

Page 24: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S24

13C-NMR 2-Naphthyl 1-thio-α-L-arabinopyranoside (23)

3040

5060

7080

90100

110

120

130

140

150

160

170

180

190

200

210

ppm

69.51269.56471.64574.958

90.222

127.121127.542128.439128.680129.313129.979130.866133.595133.783135.131

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

6271

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.9

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

9

000

SOLV

ENT

M

eOD

TD

480

74PU

LPRO

G

zgd

c30

PRO

BHD

5

mm

PA

BBO

BB−

INST

RUM

s

pect

Tim

e

2

2.43

Dat

e_

201

4091

3PR

OCN

O

1EX

PNO

3

NA

ME

K

T−D

086

OHO

OHS

OH

23

Page 25: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S25

1H-NMR α-L-Arabinopyranoside 2-naphthyl sulfone (24)

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

10.0

10.5

ppm

3.4063.4193.4283.4713.5003.6213.6983.7033.7283.7333.7593.7813.7953.8033.8174.4334.4554.6364.6414.9584.9735.2985.313

7.6767.6937.7117.7137.7307.7327.7507.7677.8537.8577.8747.8788.0658.0858.1268.1488.1938.2138.540

1.1021.0561.0251.0411.030

0.9930.896

0.886

0.896

1.0221.0371.0151.0421.0191.012

1.000PC

4

.00

GB

0LB

0

.30

Hz

SSB

0W

DW

EMSF

4

00.1

3000

36 M

Hz

SI

1310

72P1

9

.80

usec

NU

C1

1H

SFO

1

4

00.1

3320

10 M

Hz

====

====

CH

AN

NEL

f1 =

====

===

TD0

1D

1

0.0

0100

000

sec

TE

295

.7 K

DE

6

.50

usec

DW

69

.333

use

cRG

161

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

32

SOLV

ENT

D

MSO

TD

576

90PU

LPRO

G

zg

30PR

OB

HD

5

mm

PA

BB

O B

B−

INST

RUM

s

pect

Tim

e

1

4.55

Dat

e_

201

4092

3PR

OCN

O

1EX

PNO

2

NA

ME

KT−

D09

6.D

MSO

OHO

OHS

OO

OH

24

Page 26: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S26

13C-NMR α-L-Arabinopyranoside 2-naphthyl sulfone (24)

3040

5060

7080

90100

110

120

130

140

150

160

170

180

190

200

210

ppm

66.70867.73270.50273.182

92.597

124.057127.553127.868128.745129.319129.517130.575131.595134.848134.953

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

8143

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.9

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

9

000

SOLV

ENT

D

MSO

TD

480

74PU

LPRO

G

zgd

c30

PRO

BHD

5

mm

PA

BBO

BB−

INST

RUM

s

pect

Tim

e

8

.00

Dat

e_

201

4092

4PR

OCN

O

1EX

PNO

4

NA

ME

KT−

D09

6.D

MSO

OHO

OHS

OO

OH

24

Page 27: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S27

1H-NMR (2-Naphthyl)-methyl α-L-arabinopyranoside (25)

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

ppm

3.5153.5243.5383.5463.5503.5543.5813.5853.6403.6573.6623.6793.8153.8193.8233.8273.8313.8353.9043.9123.9353.9434.3424.3594.7774.8074.8895.0085.039

7.4387.4507.4527.4557.4617.4647.4707.4737.4757.4877.5307.5347.5517.5557.8237.8297.8357.8457.8497.881

1.0420.9830.9721.0091.002

1.000

1.091

1.014

1.9630.972

3.0190.954

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0078

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.1 K

DE

6

.50

usec

DW

69

.333

use

cRG

181

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

32

SOLV

ENT

M

eOD

TD

576

90PU

LPRO

G

zg

3035

2%+'�����PP�3$%%2�%%ï

INST

RUM

s

pect

Tim

e

9

.48

Dat

e_

201

4090

1PR

OCN

O

1EX

PNO

1

1$0(������������.

7ï'���

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OHO

OH

25

Page 28: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S28

13C-NMR (2-Naphthyl)-methyl α-L-arabinopyranoside (25)

3040

5060

7080

90100

110

120

130

140

150

160

170

180

190

200

210

ppm

66.93869.66171.62372.50774.251

103.911

126.930127.089127.126127.739128.657128.913128.970134.506134.767136.648

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

6271

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.8

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

9

000

SOLV

ENT

M

eOD

TD

480

74PU

LPRO

G

zgd

c30

PRO

BHD

5

mm

PA

BBO

BB−

INST

RUM

s

pect

Tim

e

2

2.59

Dat

e_

201

4090

1PR

OCN

O

1EX

PNO

3

NA

ME

K

T−D

066

OHO

OHO

OH

25

Page 29: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S29

1H-NMR 5-(2-Naphthoxy)-3-oxo-1-pentyl α-L-arabinopyranoside (26)

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

10.0

10.5

ppm3.495

3.5043.5093.5133.5183.5263.5403.5443.5603.5773.5823.5993.7323.7423.7603.7673.7773.7813.7833.7883.7923.7963.8023.8073.8403.8473.8713.8783.9213.9303.9333.9373.9453.9623.9703.9763.9833.9883.9984.2384.2414.2504.2554.2644.8887.1477.1537.1697.1757.2407.2477.2897.2927.3067.3097.3127.3277.3307.3927.3957.4097.4127.4157.4337.7377.7437.7467.758

2.0170.9854.1011.0712.0301.0333.000

0.9730.9780.9630.989

2.948

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0078

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.1 K

DE

6

.50

usec

DW

69

.333

use

cRG

181

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

32

SOLV

ENT

M

eOD

TD

576

90PU

LPRO

G

zg

30PR

OB

HD

5

mm

PA

BB

O B

B−

INST

RUM

s

pect

Tim

e

9

.54

Dat

e_

201

4090

1PR

OCN

O

1EX

PNO

1

NA

ME

K

T−D

068

OHO

OH

OH

OO

O

26

Page 30: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S30

13C-NMR 5-(2-Naphthoxy)-3-oxo-1-pentyl α-L-arabinopyranoside (26)

3040

5060

7080

90100

110

120

130

140

150

160

170

180

190

200

210

ppm

66.93468.53269.44069.64170.85071.69372.39874.188

104.986107.733

119.847124.660127.340127.859128.573130.382130.560136.128

158.144

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

6274

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.5

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

9

000

SOLV

ENT

M

eOD

TD

480

74PU

LPRO

G

zgd

c30

PRO

BHD

5

mm

PA

BBO

BB−

INST

RUM

s

pect

Tim

e

2

.32

Dat

e_

201

4090

2PR

OCN

O

1EX

PNO

3

NA

ME

K

T−D

068

OHO

OH

OH

OO

O

26

Page 31: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S31

1H-NMR 1-Naphthyl 2,3,4-tri-O-acetyl-β-D-xylopyranoside (29)

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

ppm

2.0842.1052.147

3.5793.5983.6093.6284.2634.2744.2934.3055.0425.0535.0605.0725.0795.0915.2885.3075.3255.3595.3755.3935.408

7.0657.0847.3627.3837.4027.4647.4777.4837.4927.5027.5077.5207.5417.5617.8057.8157.8227.8298.1258.1328.1408.149

5.8813.174

1.019

1.012

1.000

1.0202.067

0.983

1.0193.105

0.989

1.007

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0101

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.8 K

DE

6

.50

usec

DW

69

.333

use

cRG

161

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

64

SOLV

ENT

C

DCl

3TD

5

7690

PULP

ROG

zg30

PRO

BH

D

5 m

m P

AB

BO

BB

−IN

STRU

M

spe

ctTi

me

12.

42D

ate_

2

0140

820

PRO

CNO

1

EXPN

O

1N

AM

E

SM−D

115_

B

OAcOAcO

OAcO

29

Page 32: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S32

13C-NMR 1-Naphthyl 2,3,4-tri-O-acetyl-β-D-xylopyranoside (29)

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200

210

ppm

20.927

20.950

20.960

61.975

68.558

70.126

70.613

98.769

108.892

121.761

122.840

125.778

125.876

125.957

126.721

127.736

134.647

152.497

169.689

169.996

170.046

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

7558

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

95.2

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

3

000

SOLV

ENT

C

DCl

3TD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B−IN

STRU

M

spe

ctTi

me

0.3

1D

ate_

2

0140

821

PRO

CNO

1

EXPN

O

3N

AM

E

SM−D

115_

B

OAcOAcO

OAcO

29

Page 33: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S33

1H-NMR 4-Phenylphenyl 2,3,4-tri-O-acetyl-β-D-xylopyranoside (30)

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

ppm

3.5263.5463.5573.5764.2264.2374.2564.2685.0165.0285.0355.0475.0545.0665.2135.2205.2275.2355.2435.2515.2585.2705.2775.2845.2917.0607.0677.0727.0857.0907.0977.3027.3207.3397.4007.4207.4387.5117.5187.5237.5317.5327.5407.5507.553

9.148

1.000

1.000

1.035

3.027

2.034

1.0562.0534.045

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0100

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.7 K

DE

6

.50

usec

DW

69

.333

use

cRG

36

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

8

SOLV

ENT

C

DCl

3TD

5

7690

PULP

ROG

zg30

PRO

BH

D

5 m

m P

AB

BO

BB

−IN

STRU

M

spe

ctTi

me

8.0

4D

ate_

2

0140

909

PRO

CNO

1

EXPN

O

1N

AM

E

SM−D

123_

C

OAcOAcO

OAcO

30

Page 34: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S34

13C-NMR 4-Phenylphenyl 2,3,4-tri-O-acetyl-β-D-xylopyranoside (30)

210

200

190

180

170

160

150

140

130

120

110

100

90

80

70

60

50

40

30

20

10

ppm

20.739

20.775

20.795

61.919

68.533

70.197

70.753

98.547

117.137

126.896

127.073

128.318

128.814

136.237

140.470

156.105

169.420

169.861

169.985

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

7688

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

95.1

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

5

12SO

LVEN

T

CD

Cl3

TD

480

74PU

LPRO

G

zgd

c30

PRO

BHD

5

mm

PA

BBO

BB−

INST

RUM

s

pect

Tim

e

8

.58

Dat

e_

201

4090

9PR

OCN

O

1EX

PNO

2

NA

ME

SM

−D12

3_C

OAcOAcO

OAcO

30

Page 35: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S35

1H-NMR 2-(6-Cyano-naphthyl) 2,3,4-tri-O-acetyl-β-D-xylopyranoside (31)

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

ppm

2.0972.1002.112

3.6153.6323.6463.6634.2474.2584.2774.2885.0105.0165.0275.0335.0445.2065.2205.2245.2375.2385.2465.2645.2825.3945.407

7.2607.2627.2687.2847.2917.3717.3777.5587.5627.5797.5837.7807.8027.8137.8368.154

9.363

1.045

1.030

1.071

2.0691.068

1.1661.0371.032

2.082

1.000

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0100

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.6 K

DE

6

.50

usec

DW

69

.333

use

cRG

7

1.8

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

8

SOLV

ENT

C

DCl

3TD

5

7690

PULP

ROG

zg30

PRO

BH

D

5 m

m P

AB

BO

BB

−IN

STRU

M

spe

ctTi

me

15.

05D

ate_

2

0140

903

PRO

CNO

1

EXPN

O

1N

AM

E

SM−D

127_

A

OAcOAcO

OAcO

CN

31

Page 36: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S36

13C-NMR 2-(6-Cyano-naphthyl) 2,3,4-tri-O-acetyl-β-D-xylopyranoside (31)

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200

210

ppm

20.815

20.858

20.894

61.858

68.298

69.749

70.145

97.875

107.972

110.949

119.379

120.624

127.274

128.415

128.741

130.452

133.914

135.967

156.570

169.469

169.914

169.941

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

7611

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.6

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

3

000

SOLV

ENT

C

DCl

3TD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B−IN

STRU

M

spe

ctTi

me

0.4

0D

ate_

2

0140

904

PRO

CNO

1

EXPN

O

3N

AM

E

SM−D

127_

A

OAcOAcO

OAcO

CN

31

Page 37: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S37

1H-NMR 2-Naphthyl 2,3,4-tri-O-acetyl-1-thio-β-D-xylopyranoside (32)

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

ppm

2.0462.0572.122

3.4263.4473.4553.4774.2954.3074.3244.3374.9034.9104.9154.9244.9314.9364.9454.9574.9754.9955.0155.1815.2015.221

7.4727.4847.4857.4897.4957.4977.5037.5077.5097.5247.5287.5457.5507.7797.7937.8017.8097.8197.8327.9777.981

6.1003.058

1.014

1.007

2.0270.9871.003

2.992

3.0031.000

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0100

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

293

.6 K

DE

6

.50

usec

DW

69

.333

use

cRG

7

1.8

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

8

SOLV

ENT

C

DCl

3TD

5

7690

PULP

ROG

zg30

PRO

BH

D

5 m

m P

AB

BO

BB

−IN

STRU

M

spe

ctTi

me

7.3

5D

ate_

2

0140

826

PRO

CNO

1

EXPN

O

1N

AM

E

SM−D

121_

C

OAcOAcO

OAc

S

32

Page 38: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S38

13C-NMR 2-Naphthyl 2,3,4-tri-O-acetyl-1-thio-β-D-xylopyranoside (32)

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200

210

ppm

20.859

20.877

20.968

65.235

68.480

70.007

71.946

86.427

126.758

126.786

127.799

127.841

128.768

129.644

129.884

132.156

132.872

133.609

169.498

169.906

170.023

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

7582

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.0

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

1

000

SOLV

ENT

C

DCl

3TD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B−IN

STRU

M

spe

ctTi

me

8.0

2D

ate_

2

0140

826

PRO

CNO

1

EXPN

O

2N

AM

E

SM−D

121_

C

OAcOAcO

OAc

S

32

Page 39: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S39

1H-NMR 2-Naphthyl 2,3,4-tri-O-acetyl β-D-xylopyranosyl sulfone (33)

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

9.5

10.0

10.5

ppm

1.9922.0102.129

3.3413.3653.3703.3944.1924.2054.2214.2344.5434.5664.7684.7814.7914.8044.8144.8285.2245.2465.2685.2875.2995.310

7.6367.6397.6567.6737.6777.6967.6997.7177.7207.7347.7377.8627.8667.8837.8887.9507.9708.0028.0248.0378.0578.0678.499

3.0223.0472.926

1.000

1.009

0.982

0.989

1.0131.325

0.9801.0030.9721.0950.9791.131

0.961

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0101

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.3 K

DE

6

.50

usec

DW

69

.333

use

cRG

203

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

32

SOLV

ENT

C

DCl

3TD

5

7690

PULP

ROG

zg30

PRO

BH

D

5 m

m P

AB

BO

BB

−IN

STRU

M

spe

ctTi

me

9.2

3D

ate_

2

0140

911

PRO

CNO

1

EXPN

O

1N

AM

E

KT−

D08

8

OAcOAcO

OAc

S

OO

33

Page 40: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S40

13C-NMR 2-Naphthyl 2,3,4-tri-O-acetyl β-D-xylopyranosyl sulfone (33)

210

200

190

180

170

160

150

140

130

120

110

100

90

80

70

60

50

40

30

20

10

ppm

20.745

20.885

66.867

67.019

68.254

72.710

89.973

124.541

127.841

128.119

129.336

129.911

129.942

132.002

132.148

132.771

135.944

169.622

169.784

170.153

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

7553

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.6

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

4

500

SOLV

ENT

C

DCl

3TD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B−IN

STRU

M

spe

ctTi

me

4.4

5D

ate_

2

0140

912

PRO

CNO

1

EXPN

O

3N

AM

E

KT−

D08

8

OAcOAcO

OAc

S

OO

33

Page 41: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S41

1H-NMR 2-Naphthyl 2,3,4-tri-O-acetyl β-L-xylopyranoside (35)

13

12

11

10

98

76

54

32

1ppm

2.1002.1052.1133.5793.5983.6093.6284.2594.2704.2894.3015.0205.0315.0385.0505.0565.0695.2205.2345.2385.2515.2545.2565.2735.2925.3315.3467.1677.1737.1897.1957.3637.3687.3857.3887.3917.4057.4087.4417.4447.4587.4617.4647.4787.4827.7337.7537.7687.7807.7907.800

8.94

1.00

1.01

1.012.021.03

0.952.021.053.05

PC 4.00

GB 0

LB 0.30 Hz

SSB 0

WDW EM

SF 400.1300098 MHz

SI 131072

SFO1 400.1334011 MHz

PL1W 27.00716019 W

PL1

−5

.00

dBP1 9.10 usec

NUC1 1H

======== CHANNEL f1 ========

TD0 1

D1 0.00100000 sec

TE 300.0 K

DE 6.00 usec

DW 69.333 usec

RG 456

AQ 3.9998901 sec

FIDRES 0.125005 Hz

SWH 7211.539 Hz

DS 2

NS 32

SOLVENT CDCl3

TD 57690

PULPROG zg30

PROBHD 5 mm PABBO BB/

INSTRUM spect

Time 8.19

Date_ 20130809

PROCNO 1

EXPNO 1

NAME

K

H−B2

00b

OAcO

AcO

OOAc

35

Page 42: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S42

13C-NMR 2-Naphthyl 2,3,4-tri-O-acetyl β-L-xylopyranoside (35)

210

200

190

180

170

160

150

140

130

120

110

100

90

80

70

60

50

40

30

20

10

ppm

20.899

20.931

20.956

62.011

68.636

70.240

70.709

98.669

111.443

118.936

124.772

126.742

127.302

127.828

129.826

130.175

134.295

154.464

169.574

170.014

170.097

SF

100

.612

7546

MH

zSI

3

2768

SFO

2

4

00.1

3160

05 M

Hz

PL12

W

0

.349

5255

4 W

PL2W

27.

0071

6019

WPL

12

1

3.88

dB

PL2

−5.

00 d

BPC

PD2

80.0

0 us

ecN

UC2

1

HCP

DPR

G2

wal

tz16

====

====

CH

AN

NEL

f2 =

====

===

SFO

1

1

00.6

2282

98 M

Hz

PL1W

73.

8220

0623

WPL

1

4.00

dB

P1

8.0

0 us

ecN

UC1

1

3C==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

3

00.0

KD

E

6.0

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

8

000

SOLV

ENT

C

DCl

3TD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B/IN

STRU

M

spe

ctTi

me

22.

33D

ate_

2

0130

809

PRO

CNO

1

EXPN

O

3N

AM

E

KH

−B20

0b

OAcO

AcO

OOAc

35

Page 43: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S43

1H-NMR 2-Naphthyl 2,3,4-tri-O-acetyl 1-thio-α-L-arabinopyranoside (38)

109

87

65

43

21

ppm

2.0682.0862.128

3.6873.6923.7193.7244.1834.1944.2144.2254.9154.9345.1205.1295.1415.1495.2825.2885.2935.2985.3025.3075.3227.4667.4787.4817.4917.5017.5047.5167.5517.5567.5737.5777.7797.7907.7947.8017.8107.8157.8298.0038.008

3.0252.9533.061

1.000

1.023

0.9701.0072.016

2.0251.0243.0780.974

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0101

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.8 K

DE

6

.50

usec

DW

69

.333

use

cRG

181

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

32

SOLV

ENT

C

DCl

3TD

5

7690

PULP

ROG

zg30

PRO

BH

D

5 m

m P

AB

BO

BB

−IN

STRU

M

spe

ctTi

me

9.0

8D

ate_

2

0140

909

PRO

CNO

1

EXPN

O

1N

AM

E

KT−

D08

0

O

AcO

OAc

S

OAc

38

Page 44: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S44

13C-NMR 2-Naphthyl 2,3,4-tri-O-acetyl 1-thio-α-L-arabinopyranoside (38)

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200

210

ppm

20.879

21.036

65.441

67.629

68.733

70.581

87.061

126.676

126.777

127.756

127.850

128.738

129.622

130.619

131.693

132.814

133.647

169.592

170.108

170.387

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

7561

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

95.1

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

9

000

SOLV

ENT

C

DCl

3TD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B−IN

STRU

M

spe

ctTi

me

22.

44D

ate_

2

0140

909

PRO

CNO

1

EXPN

O

3N

AM

E

KT−

D08

0

O

AcO

OAc

S

OAc

38

Page 45: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S45

1H-NMR 2-Naphthyl 2,3,4-tri-O-acetyl α-L-arabinopyranosyl sulfone (39)

109

87

65

43

21

ppm

1.637

1.9912.151

3.6623.6643.6943.6974.0764.0824.1094.1154.1274.5374.5605.0405.0485.0645.0725.1305.1375.1415.5035.5275.5507.6247.6277.6417.6447.6617.6657.6787.6827.6997.7027.7167.7197.9257.9307.9477.9517.9998.0218.0538.0738.557

3.171

3.2213.275

1.069

1.303

1.031

1.0801.048

1.029

2.1422.0681.1101.142

1.000

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0101

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.6 K

DE

6

.50

usec

DW

69

.333

use

cRG

181

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

32

SOLV

ENT

C

DCl

3TD

5

7690

PULP

ROG

zg30

PRO

BH

D

5 m

m P

AB

BO

BB

−IN

STRU

M

spe

ctTi

me

8.3

8D

ate_

2

0140

918

PRO

CNO

1

EXPN

O

1N

AM

E

KT−

D09

2

O

AcO

OAc

S

OO

OAc

39

Page 46: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S46

13C-NMR 2-Naphthyl 2,3,4-tri-O-acetyl α-L-arabinopyranosyl sulfone (39)

210

200

190

180

170

160

150

140

130

120

110

100

90

80

70

60

50

40

30

20

10

ppm

20.396

20.754

20.997

64.581

67.583

67.723

70.962

90.080

124.867

127.739

127.984

128.903

129.809

129.918

132.070

132.194

132.827

135.823

169.629

170.057

170.156

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

7555

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

95.4

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

3

072

SOLV

ENT

C

DCl

3TD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B−IN

STRU

M

spe

ctTi

me

21.

43D

ate_

2

0140

918

PRO

CNO

1

EXPN

O

3N

AM

E

KT−

D09

2

O

AcO

OAc

S

OO

OAc

39

Page 47: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S47

1H-NMR (2-Naphthyl)-methyl 2,3,4-tri-O-acetyl α-L-arabinopyranoside (40)

12

11

10

98

76

54

32

1ppm

2.0022.0312.1273.5903.5943.6223.6264.0394.0474.0714.0804.5094.5264.7324.7634.9945.0035.0165.0265.0455.2495.2545.2585.2625.2665.2745.2915.2975.3147.3937.3977.4147.4187.4447.4577.4597.4617.4697.4807.4827.4947.7457.7927.8067.8167.827

2.923.073.05

1.00

1.10

1.011.02

2.022.08

1.022.051.033.09

PC 4.00

GB 0

LB 0.30 Hz

SSB 0

WDW EM

SF 400.1300095 MHz

SI 131072

P1 9.80 usec

NUC1 1H

SFO1 400.1332010 MHz

======== CHANNEL f1 ========

TD0 1

D1 0.00100000 sec

TE 294.1 K

DE 6.50 usec

DW 69.333 usec

RG 25.4

AQ 3.9998901 sec

FIDRES 0.125005 Hz

SWH 7211.539 Hz

DS 2

NS 32

SOLVENT CDCl3

TD 57690

PULPROG zg30

PROB

HD

5 m

m PA

BBO

BB−

INSTRUM spect

Time 16.37

Date_ 20140825

PROCNO 1

EXPNO 1

NAME

K

T−D0

58d

O

AcO

OAcO

OAc

40

Page 48: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S48

13C-NMR (2-Naphthyl)-methyl 2,3,4-tri-O-acetyl α-L-arabinopyranoside (40)

210

200

190

180

170

160

150

140

130

120

110

100

90

80

70

60

50

40

30

20

10

ppm

14.116

20.652

20.782

20.898

22.665

31.846

63.039

67.603

69.176

70.102

70.363

99.433

125.481

126.066

126.253

126.507

127.710

127.826

128.209

132.995

133.143

134.429

169.417

170.066

170.289

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

7729

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.5

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

9

000

SOLV

ENT

C

DCl

3TD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B−IN

STRU

M

spe

ctTi

me

0.4

1D

ate_

2

0140

826

PRO

CNO

1

EXPN

O

3N

AM

E

KT−

D05

8d

O

AcO

OAcO

OAc

40

Page 49: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S49

1H-NMR 5-Chloro-3-oxo-1-pentyl 2,3,4-tri-O-acetyl-α-L-arabinopyranoside (42)

1.5

2.0

2.5

3.0

3.5

4.0

4.5

5.0

5.5

6.0

6.5

7.0

7.5

8.0

8.5

9.0

ppm

2.0722.1323.5973.5993.6123.6153.6213.6273.6503.6543.6653.6723.6823.6843.6913.6983.7053.7263.7333.7403.7493.7533.7563.9263.9353.9443.9513.9583.9633.9714.0094.0184.0424.0504.5004.5175.0195.0285.0435.0515.1735.1905.1975.2145.2485.2525.2565.2615.2655.269

3.223.123.30

8.30

1.031.03

1.00

1.021.021.02

PC 4.00

GB 0

LB 0.30 Hz

SSB 0

WDW EM

SF 400.1300101 MHz

SI 131072

P1 9.80 usec

NUC1 1H

SFO1 400.1332010 MHz

======== CHANNEL f1 ========

TD0 1

D1 0.00100000 sec

TE 293.8 K

DE 6.50 usec

DW 69.333 usec

RG 161

AQ 3.9998901 sec

FIDRES 0.125005 Hz

SWH 7211.539 Hz

DS 2

NS 32

SOLVENT CDCl3

TD 57690

PULPROG zg30

PROB

HD

5 m

m PA

BBO

BB−

INSTRUM spect

Time 10.54

Date_ 20140825

PROCNO 1

EXPNO 1

NAME

K

T−D0

56f

O

AcO

OAc

OAc

OO

Cl

42

Page 50: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S50

13C-NMR 5-Chloro-3-oxo-1-pentyl 2,3,4-tri-O-acetyl-α-L-arabinopyranoside (42)

210

200

190

180

170

160

150

140

130

120

110

100

90

80

70

60

50

40

30

20

10

ppm

20.830

20.963

21.078

42.975

63.332

67.797

68.826

69.246

70.314

70.507

71.550

101.132

169.668

170.316

170.479PC

1

.40

GB

0LB

1

.00

Hz

SSB

0W

DW

EMSF

1

00.6

1275

64 M

Hz

SI

327

68P1

9

.80

usec

NU

C1

13C

SFO

1

1

00.6

2282

98 M

Hz

====

====

CH

AN

NEL

f1 =

====

===

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

94.9

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

9

000

SOLV

ENT

C

DCl

3TD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B−IN

STRU

M

spe

ctTi

me

23.

43D

ate_

2

0140

822

PRO

CNO

1

EXPN

O

3N

AM

E

KT−

D05

6c

O

AcO

OAc

OAc

OO

Cl

42

Page 51: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S51

1H-NMR 5-(2-Naphthoxy)-3-oxo-1-pentyl 2,3,4-tri-O-acetyl-α-L-arabinopyranoside (43)

109

87

65

43

21

ppm2.014

2.0412.1133.5603.5643.5923.5973.7263.7323.7363.7433.7523.7603.8833.8843.8953.8993.9083.9613.9703.9763.9853.9883.9934.0174.0264.2094.2164.2214.2334.4884.5055.0085.0175.0315.0405.1855.2025.2085.2265.2305.2345.2395.2435.2477.1347.1407.1587.1647.1807.1867.3097.3237.3267.3297.3437.3467.4077.4107.4247.4277.4317.4457.4487.7087.7237.7287.745

3.0822.9592.916

1.0423.0512.0312.0751.992

0.991

1.0591.0060.981

0.9920.9751.0061.0002.979

PC

4.0

0G

B

0

LB

0.3

0 H

zSS

B

0

WD

W

EM

SF

400

.130

0099

MH

zSI

13

1072

P1

9.8

0 us

ecN

UC1

1

HSF

O1

400

.133

2010

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

1

0.0

0100

000

sec

TE

294

.7 K

DE

6

.50

usec

DW

69

.333

use

cRG

4

0.3

AQ

3.99

9890

1 se

cFI

DRE

S

0

.125

005

Hz

SWH

7211

.539

Hz

DS

2N

S

32

SOLV

ENT

C

DCl

3TD

5

7690

PULP

ROG

zg30

PRO

BH

D

5 m

m P

AB

BO

BB

−IN

STRU

M

spe

ctTi

me

15.

15D

ate_

2

0140

827

PRO

CNO

1

EXPN

O

1N

AM

E

KT−

D06

0

O

AcO

OAc

OAc

OO

O

43

Page 52: Supporting - Royal Society of Chemistry · S1 Supporting Information Exploration of the active site of β4GalT7: Modifications of the aglycon of aromatic xylosides Anna Siegbahn,a#

  S52

13C-NMR 5-(2-Naphthoxy)-3-oxo-1-pentyl 2,3,4-tri-O-acetyl-α-L-arabinopyranoside (43)

 

210

200

190

180

170

160

150

140

130

120

110

100

90

80

70

60

50

40

30

20

10

ppm

20.741

20.841

20.980

63.186

67.477

67.733

68.868

69.192

69.906

70.240

70.629

101.102

106.749

119.010

123.754

126.441

126.807

127.703

129.083

129.492

134.529

156.756

169.624

170.218

170.388

PC

1.4

0G

B

0

LB

1.0

0 H

zSS

B

0

WD

W

EM

SF

100

.612

7649

MH

zSI

3

2768

P1

9.8

0 us

ecN

UC1

1

3CSF

O1

100

.622

8298

MH

z==

====

== C

HA

NN

EL f1

===

====

=

TD0

1D

11

0.0

3000

000

sec

D1

0

.001

0000

0 se

cTE

2

95.0

KD

E

6.5

0 us

ecD

W

20.8

00 u

sec

RG

5

7A

Q

0.

9999

892

sec

FID

RES

0.5

0003

0 H

zSW

H

240

38.4

61 H

zD

S

4

NS

9

000

SOLV

ENT

C

DCl

3TD

4

8074

PULP

ROG

z

gdc3

0PR

OBH

D

5 m

m P

ABB

O B

B−IN

STRU

M

spe

ctTi

me

22.

45D

ate_

2

0140

827

PRO

CNO

1

EXPN

O

3N

AM

E

KT−

D06

0

O

AcO

OAc

OAc

OO

O

43