supplementary data 1: ms and ms/ms data for metabolite identification
DESCRIPTION
Table S1: Summary of identified metabolites Identified metabolitesMALDI in situ MS/MSLC-MS full scanLC-MS/MSIdentified by accurate MS (accuracy, 2ppm) AMPyes - ADPyes - ATPyes - UMPyes -- UDPyes -- UTPyes -- GMPyes -- GDPyes -- GTPyes -- Glucose mono phosphate-yes-- Fructose-1,6-bis-phosphate-yes-- N-acetylaspartete-yes-- Citrate-yes - IMP-yes-- Glutathione-yes - Glycerol-mono-phosphate-yes-- Desulfoglucotropeolin---yes Pyrophosphate---yes Sn-glycerol-3-Phosphoethanolamine---yes 3-Sulfino-L-alanine---yes Three Glycerophosphoinositols---yesTRANSCRIPT
Supplementary data 1:MS and MS/MS data for metabolite identification
The identification of metabolites were carried out by in situ MS/MS using MALDI-FTICR and LC-MS/MS using Waters Acquity UPLC coupled to maXis UHR-TOF-MS. Table S1 summarized the 23 identified metabolite.
Table S1: Summary of identified metabolitesIdentified metabolites MALDI in situ MS/MS LC-MS full scan LC-MS/MS Identified by accurate
MS (accuracy, 2ppm)
AMP yes yes yes -
ADP yes yes yes -
ATP yes yes yes -
UMP yes yes - -
UDP yes yes - -
UTP yes yes - -
GMP yes yes - -
GDP yes yes - -
GTP yes yes - -
Glucose mono phosphate - yes - -
Fructose-1,6-bis-phosphate - yes - -
N-acetylaspartete - yes - -
Citrate - yes yes -
IMP - yes - -
Glutathione - yes yes -
Glycerol-mono-phosphate - yes - -
Desulfoglucotropeolin - - - yes
Pyrophosphate - - - yes
Sn-glycerol-3-Phosphoethanolamine - - - yes
3-Sulfino-L-alanine - - - yes
Three Glycerophosphoinositols - - - yes
LC-MS/MS data
Standards used for the LC-MS/MS experiments
Name Sigma Name Empirical FormulaSigma Catalog No.
AMP Adenosine 5 -monophosphate monohydrate′ C10H14N5O7P · H2O A2252-5GADP Adenosine 5 -diphosphate sodium salt′ C10H15N5O10P2 A2754-100MG
ATPAdenosine 5 -triphosphate (ATP) disodium salt ′hydrate C10H14N5O13P3Na2 · xH2O FLAAS-1VL
UMP Uridine 5 -monophosphate disodium salt′ C9H13N2O9P U6375-1GUDP Uridine 5 -diphosphate disodium salt hydrate′ C9H12N2Na2O12P2 · xH2O 94330-100MGUTP Uridine 5 -triphosphate trisodium salt dihydrate′ C9H12N2O15P3Na3 94370-250MG
GMPGuanosine 5 -monophosphate disodium salt ′hydrate C10H12N5Na2O8P · xH2O G8377-500MG
GDP Guanosine 5 -diphosphate sodium salt′ C10H15N5O11P2 G7127-25MGGTP Guanosine 5 -triphosphate sodium salt hydrate′ C10H16N5O14P3 · xNa+ G8877-25MGIMP Inosine 5 -monophosphate disodium salt hydrate′ C10H11N4O8PNa2 · xH2O I4625-5G
Glutathione L-Glutathione reducedH2NCH(CO2H)CH2CH2CONHCH(CH2SH)CONHCH2CO2H G4251-1G
Glucose monophosphateD-Glucose 6-phosphate disodium salt hydrate Sigma Grade, ≥98% C6H11Na2O9P · xH2O G7250-500MG
Fructose 1,6-bisphosphate
D-Fructose 1,6-bisphosphate trisodium salt hydrate C6H11Na3O12P2 · xH2O F6803-1G
Glycerol monophosphate sn-Glycerol 3-phosphate lithium salt C3H9O6P · xLi+ 94124-10MGN-Acetylaspartate N-Acetyl-L-aspartic acid HO2CCH2CH(NHCOCH3)CO2H 00920-5GCitrate Citric acid HOC(COOH)(CH2COOH)2 251275-5G
10.4 Time [min]0.0
0.2
0.4
0.6
0.84x10
Intens.
10.4 Time [min]0
1
2
4x10Intens.
AMP
10.4 Time [min]0.0
0.5
1.0
4x10Intens.
10.4 Time [min]0.00
0.25
0.50
0.75
1.00
4x10Intens.
ADP
10.4 Time [min]0
2000
4000
6000
Intens.
10.4 Time [min]0
2000
4000
6000
Intens.
ATPst
anda
rdRe
tina
tissu
eGMP GDP GTP
UMP
10.4 Time [min]0
2000
4000
6000
Intens.
10.4 Time [min]0
1000
2000
3000Intens.
UDP
10.5 Time [min]0.0
0.5
1.0
1.5
4x10Intens.
10.5 Time [min]0
1000
2000
Intens.
UTP Glu-6-P Fru-1,6-P
10.5 Time [min]0.00
0.25
0.50
0.75
1.00
4x10Intens.
10.5 Time [min]0
1000
2000
3000
Intens.
10.5 Time [min]0
500
1000
1500
Intens.
10.5 Time [min]0
2
4
6
4x10Intens.
10.5 Time [min]0.0
0.5
1.0
4x10Intens.
10.5 Time [min]0
50
100
150
200
Intens.
10.5 Time [min]0.0
0.2
0.4
0.6
0.8
4x10Intens.
10.5 Time [min]0
200
400
600
800
Intens.
10.5 Time [min]0
1000
2000
3000
Intens.
10.5 Time [min]0.0
0.5
1.0
5x10Intens.
10.5 Time [min]0
2
4
4x10Intens.
10.5 Time [min]0
500
1000
1500
Intens.
stan
dard
Retin
a tis
sue
Full scan LC-MS data
stan
dard
Retin
a tis
sue
Glycerol-3-P
10.4 Time [min]0.0
0.5
1.0
4x10Intens.
10.4 Time [min]0.0
0.5
1.0
4x10Intens.
N-Asp-Ac Glutathione Citrate IMP
10.4 Time [min]0.0
0.5
1.0
4x10Intens.
10.4 Time [min]0
1000
2000
3000
Intens.
10 Time [min]0
2
4
5x10Intens.
10 Time [min]0.0
0.5
1.0
5x10Intens.
10 Time [min]0
1
2
4x10Intens.
10 Time [min]0.00
0.25
0.50
0.75
1.005x10
Intens.
10 Time [min]0.0
0.5
1.0
1.5
5x10Intens.
10 Time [min]0
2
4
6
4x10Intens.
Full scan LC-MS data
AMP ADP ATP
stan
dard
Retin
a tis
sue
LC-MS/MS data
174.0185MS2(346.0000), 10.3-10.4min
0.0
0.5
1.0
1.5
2.0
4x10Intens.
100 200 300 400 m/z
174.0183
283.2279
MS2(346.0093), 10.4-10.5min
0
200
400
600
800
1000
1200
Intens.
100 200 300 400 m/z
158.9051
212.9347
272.9215407.9591
MS2(425.9661), 10.5-10.6min
0
1000
2000
3000
4000
5000
6000Intens.
100 200 300 400 m/z
158.9049
272.9208
MS2(425.9671), 10.5-10.6min
0
500
1000
1500
Intens.
100 200 300 400 m/z
158.9050
272.9223
407.9585
MS2(505.9238), 10.6-10.7min
0
2000
4000
6000
Intens.
100 200 300 400 m/z
158.9049
238.8591
407.9587
MS2(505.9237), 10.6-10.7min
0
500
1000
1500
Intens.
100 200 300 400 m/z
Citrate Glutathione
stan
dard
Retin
a tis
sue
LC-MS/MS data
86.9986
110.9947
136.9189160.9511
178.9601
MS2(191.0000), 10.6-12.6min
0
100
200
300
400
Intens.
50 100 150 m/z
86.9973
110.9944
159.8727
178.9599
MS2(191.0000), 10.6-11.9min
0
50
100
150
200
250
300
Intens.
50 100 150 m/z
143.0276
170.9843
242.0486
MS2(306.0000), 10.4-10.6min
0
200
400
600
800
1000
Intens.
50 100 150 200 250 m/z
143.0277MS2(306.0000), 10.3-10.7min
0
500
1000
1500
Intens.
50 100 150 200 250 m/z
in situ MS/MS data
78.9590
96.9696
134.0471
150.9800
192.9905
211.0010
321.0309
346.0550
103.4405
146.5199
211.0012
320.8274
346.0547
0.00
0.25
0.50
0.75
1.00
1.25
1.50
7x10Intens.
0.0
0.5
1.0
1.5
6x10
100 150 200 250 300 350 400 m/z
AMP
Reference MS/MS
In situMS/MS
ADP
Reference MS/MS
103.4406
146.4992
272.9565
328.0445
426.0206
103.4405
146.4765
213.7404
328.0444
0
1
2
3
4
5
6x10Intens.
0.0
0.5
1.0
1.5
6x10
100 150 200 250 300 350 400 450 500 550 m/z
272.9564
In situMS/MS
ATP
Reference MS/MS
In situMS/MS
71.4259 122.9010
158.9252272.9565
321.0653
408.0102
448.0030
505.9864
72.0630
103.4406
146.4884
281.2478 321.4463
408.0103
0.00
0.25
0.50
0.75
1.00
1.25
1.50
7x10Intens.
0.0
0.5
1.0
1.5
6x10
100 150 200 250 300 350 400 450 500 m/z
UMP
Reference MS/MS
78.9591
96.9696150.9801 192.9905
211.0010
262.0117
280.0222
323.0278
103.4406
146.6373
189.4563
211.0014
265.1477
323.0278
0.0
0.5
1.0
1.5
2.0
7x10Intens.
0.0
0.5
1.0
1.5
6x10
100 150 200 250 300 350 m/z
In situMS/MS
UDP
Reference MS/MS
111.0199158.9252
201.4972272.9566
305.0173
384.8028
402.9936
103.4405
146.5863
268.9469
305.0171
321.3053
346.0549
384.8028
402.9937
0.0
0.2
0.4
0.6
0.8
1.0
1.28x10
Intens.
0.0
0.5
1.0
1.5
2.0
6x10
100 150 200 250 300 350 400 m/z
In situMS/MS
UTPReference MS/MS
158.9252
238.8915
272.9565
300.8470
384.9832
482.9592
103.4405
146.5251
272.9566 320.9261
384.9834
465.9558
0
1
2
3
7x10Intens.
0
1
2
3
4
6x10
50 100 150 200 250 300 350 400 450 500 m/z
In situMS/MS
GMP
Reference MS/MS
105.5006 146.4845
211.0010
362.0497
80.3442
103.4406
146.4955
211.0011 321.2326
362.2362
0
1
2
3
4
7x10Intens.
0.0
0.5
1.0
1.5
2.0
6x10
100 150 200 250 300 350 400 m/z
In situMS/MS
77.3744 103.4405
150.0420
172.0199 321.7295
344.0393
424.0044
GDP_MSMS_0_M9_000002.d: -MS2(qCID 442.0157)
122.9010
146.6474
321.1789
344.0390
Retina_GDP_MSMS_0_D8_000001.d: -MS2(qCID 442.0000)0.00
0.25
0.50
0.75
1.00
1.25
1.50
7x10Intens.
0.0
0.5
1.0
1.5
6x10
100 150 200 250 300 350 400 m/z
GDP
Reference MS/MS
In situMS/MS
GTP
Reference MS/MS
In situMS/MS
176.9357
212.0029
272.9566
424.0049
485.9791
521.9812
146.6842
176.9357 214.1420272.9565 328.0437
424.0049
521.9816
0.00
0.25
0.50
0.75
1.00
1.25
8x10Intens.
0
1
2
3
4
6x10
150 200 250 300 350 400 450 500 550 m/z