comparative differential cuproproteomes of rhodobacter … · 2020-02-28 · s - 1 comparative...
TRANSCRIPT
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Comparative Differential Cuproproteomes of Rhodobacter capsulatus Reveal Novel Copper Homeostasis related Proteins
Nur Selamoglua, Özlem Öndera,b, Yavuz Öztürka,e, Bahia Khalfaoui-Hassania,#, Crysten E. Blaby-
Haasc, Benjamin A. Garciad, Hans-Georg Koche and Fevzi Daldala*
aDepartment of Biology, University of Pennsylvania, Philadelphia, PA, 19104 USA; bDepartment
of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania,
Philadelphia, PA 19104, USA; cBiology Department, Brookhaven National Laboratory, Upton,
NY 11973, USA; dDepartment of Biochemistry and Biophysics, Perelman School of Medicine,
University of Pennsylvania, Philadelphia, PA 19104, USA, eInstitute of Biochemistry and
Molecular Biology, ZBMZ, Faculty of Medicine, Albert-Ludwigs University of Freiburg, 79104
Freiburg, Germany.
Running Title: Rhodobacter capsulatus Copper-Responsive Proteome
Key words: Rhodobacter capsulatus proteome, differential cuproproteomics, copper
homeostasis, copper-responsive proteins, uncharacterized proteins.
#Present address: IPREM, UMR CNRS 5254, and Université de Pau et des Pays de l’Adour,
BP1155 Pau, France
*Corresponding author: Fevzi Daldal
[email protected]; +1 215-898-4394
Files in this Data Supplement:
Supplemental Information and Table S3 (Excel file)
Electronic Supplementary Material (ESI) for Metallomics.This journal is © The Royal Society of Chemistry 2020
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SUPPLEMENTAL INFORMATION
Experimental Procedures
ProteaseMAXTM Extraction/Trypsin Digestion (protocol B). Each 100 mg aliquot from the
resulting wet pellets was washed with ultrapure water, centrifuged and the supernatant discarded.
Solubilization and extraction of proteins was performed using ProteaseMAXTM Surfactant Trypsin
Enhancer (Promega Inc.), followed by trypsin digestion as follows (1). The pellet was resuspended
in 0.5 ml of ProteaseMAXTM solution (1 mg/ml in 10% acetonitrile and 90% 50mM NH4HCO3),
and 10 min of rigorous vortexing was followed by incubation in a 95 °C water bath for 10 min.
Following 15 min of sonication at 4 °C (BioruptorR, Diagenode), the sample was vortexed at room
temperature for 30 min. Estimates from separate BCA analysis of similar samples indicated that ~
400-500 µg protein was solubilized at this stage. Further reduction/alkylation and digestions steps
were completed without separating the insolubilized solid fraction. Proteins were reduced by
adding dithiothreitol (DTT) to a final concentration of 5mM and incubating for 30 min at 37 °C,
and alkylated by adding iodoacetamide to a final concentration of 15 mM and incubating for 30
min at room temperature in the dark. Trypsin (Promega, Sequencing Grade Modified Trypsin) was
added to the mixture at a 1/100 w/w ratio of enzyme/protein, and the extract was digested overnight
at 37 °C, pH 7. Digested mixtures were vortexed for 10 min and acidified with 10% formic acid
to pH 3-4 to halt digestion, vortexed and placed in a 45 °C shaker for 30 min to degrade
ProteaseMAXTM. Following a 15 min centrifugation at 15,000 x g, the supernatants were removed
and dried to completeness in a vacuum centrifuge at 45 °C. The digested peptide samples were
resuspended in 50 µL 5% acetonitrile/0.1% formic acid solution, and desalted using ZipTips
(Milipore U-C18 P10), eluted, lyophilized and stored at -80 °C. Samples were resuspended in 10
µL (5% acetonitrile/0.1% formic acid) prior to MS analysis.
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The efficacy of the extraction/digestion protocols A (thiourea/urea/trypsin/LysC) and B
(ProteaseMax/trypsin) was determined using the nano-LC-MS/MS Thermo Orbitrap Velos Pro
system described below. Protocol A yielded ~40% more identified proteins than protocol B (1784
versus 1244 in triplicate samples), indicating that the former with multiple digestion steps was
more effective than the latter. In addition, a further 15-20% increase in the number of identified
proteins was observed when the samples prepared by protocol A were analyzed using the higher
sensitivity Q-Exactive MS, rendering it the method of choice for the quantitative aspects of this
study. The qualitative data for the protein identification included all datasets, including those
treated by the procedure A or B, analyzed using Orbitrap Velos (c.f. below) as well as the Q-
Exactive MS runs (c.f. main text).
Data acquisition using nano-LC-MS/MS Thermo Orbitrap Velos Pro. Prepared digests were
loaded without pre-fractionation onto a 20 cm column (packed in-house, 75 µm ID, 3µm C18,
RepoSil-Pur Q18-AQ (Dr.Maisch GmbH)) with buffer A (0.1% formic acid). Peptides were
separated and eluted by nano-HPLC (EASY-nLC 1000, Thermo Inc.) using a 85 min linear
gradient from 4% to 40% buffer B (0.1% formic acid in acetonitrile), followed by a 20 min linear
gradient from 40% to 90% B, followed by 15 min washing with 90% B (flow rate 300 nL/min),
and re-equilibration with buffer A. The nLC system was coupled to an Orbitrap Velos Pro mass
spectrometer (Thermo Scientific), programmed to acquire spectra in data-dependent mode, with
high resolution full survey scans followed by the top 15 most abundant ions fragmented for MS2
(Thermo Xcalibur 2.2 software). The resolution for full MS scans (300-2000 m/z) was 60,000 (at
400 m/z) and dual lock mass was enabled with 371.101233 m/z and 445.120024 m/z polysiloxane
background ions. The 15 most intense ions were selected for fragmentation by collision induced
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dissociation (CID) with collision energy set at 35%, and with dynamic exclusion enabled to
minimize repeat fragmentation (repeat count: 3, repeat duration: 30 sec, exclusion duration: 90
sec; exclusion list: 500). Unassigned charge states and singly charged species were rejected. The
raw data were analyzed as described in the main text.
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REFERENCES
1. Pirmoradian, M., Budamgunta, H., Chingin, K., Zhang, B., Astorga-Wells, J., and Zubarev, R. A. (2013) Rapid and deep human proteome analysis by single-dimension shotgun proteomics. Mol Cell Proteomics 12, 3330-3338
2. Sambrook, J., and Russell, D. W. (2001) Molecular Cloning: a laboratory manual, 3rd ed. ed., Cold Spring harbor Laboratory Press, Cold Spring harbor
3. Simon, R., Priefer, U., and Puhler, A. (1983) A Broad Host Range Mobilization System for Invivo Genetic-Engineering - Transposon Mutagenesis in Gram-Negative Bacteria. Bio-Technology 1, 784-791
4. Zannoni, D., Prince, R. C., Dutton, P. L., and Marrs, B. L. (1980) Isolation and Characterization of a Cytochrome c2-Deficient Mutant of Rhodopseudomonas-capsulata. Febs Letters 113, 289-293
5. Yen, H. C., Hu, N. T., and Marrs, B. L. (1979) Characterization of the gene transfer agent made by an overproducer mutant of Rhodopseudomonas capsulata. J Mol Biol 131, 157-168
6. Ditta, G., Schmidhauser, T., Yakobson, E., Lu, P., Liang, X. W., Finlay, D. R., Guiney, D., and Helinski, D. R. (1985) Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression. Plasmid 13, 149-153
7. Brimacombe, C. A., Stevens, A., Jun, D., Mercer, R., Lang, A. S., and Beatty, J. T. (2013) Quorum-sensing regulation of a capsular polysaccharide receptor for the Rhodobacter capsulatus gene transfer agent (RcGTA). Mol Microbiol 87, 802-817
8. S. Ekici, S. Turkarslan, G. Pawlik, A. Dancis, N. S. Baliga, H. G. Koch and F. Daldal, Intracytoplasmic copper homeostasis controls cytochrome c oxidase production, MBio, 2014, 5, e01055-01013.
9. Y. Zhang, C. E. Blaby-Haas, S. Steimle, A. F. Verissimo, V. A. Garcia-Angulo, H. G. Koch, F. Daldal and B. Khalfaoui-Hassani, Cu Transport by the Extended Family of CcoA-like Transporters (CalT) in Proteobacteria, Sci Rep, 2019, 9, 1208.
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Table S1. Strains and plasmids used in this work. Strain or plasmid Description Phenotype Reference
Strains E. coli HB101
F- Δ(gpt-proA)62 leuB6 supE44 ara-14 galK2 lacY1 Δ(mcrC-mrr) rpsL20 (StrR) xyl-5 mtl-1recA13
Strr
(2)
S17-1 recA pro hsdR, RP4(Tc::Mu, Km::Tn7) (3)
R. capsulatus
aMT1131
crtD121
wild type (Rifr)
(4)
Y262 GTA overproducer (5)
YO-Δ00098 Δ(rcc00098) In-frame, markerless This work
YO-Δ00738 Δ(rcc00738::Gm) Gmr This work
YO- Δ00885 Δ(rcc00891-rcc00885::Gm) Gmr This work
YO-Δ01027 Δ(rcc01027-rcc01031::Gm) Gmr, This work
YO-Δ01423Su Δ(rcc01423) In-frame, markerless This work
YO-Δ01423 Δ(rcc01423::Gm) Gmr This work
YO-Δ01445 Δ(rcc01445::Gm) Gmr This work
YO-Δ02109 Δ(rcc02109::Gm) Gmr This work
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YO-Δ02111 Δ ( rcc02111) In-frame, markerless This work
YO-Δ03065 Δ(rcc03065-rcc03067::Gm) Gmr This work
Plasmids
pRK2013 Conjugation helper
Kanr
(6)
pRK415 Broad host-range vector
Tetr
(6)
pZDJ Puc promoter replaced with tetA promoter on the suicide plasmid pZJD29a
sacB, GmR (7)
pYO98 Δ(rcc01423::Gm) on pRK415 Gmr This work
pYO-Δ00738 Δ(rcc00738::Gm) on pRK415 Tetr This work
pYO-Δ00885 Δ(rcc00891-rcc00885::Gm) on pRK415 Tetr, Gmr This work
pYO-Δ01027 Δ(rcc01027-rcc01031::Gm) on pRK415 Tetr, Gmr This work
pYO-Δ01445 Δ(rcc01445::Gm) on pRK415 Tetr, Gmr This work
pYO-Δ03065 Δ(rcc03065-rcc03067::Gm) on pRK415 Tetr, Gmr This work
pYO-Δ02109 Δ(rcc02109::Gm) on pRK415 Tetr, Gmr This work
pYO97 Δ(rcc01423) on pZDJ; in-frame deletion of rcc01423 without its first 4 and last 4 codons.
Gmr This work
pYO-Δ00098 Δ(rcc00098) on pZDJ; in-frame deletion of rcc02111 without its first 4 and last 4 codons.
Gmr This work
pYO-Δ02111Su Δ(rcc02111) on pZDJ; in-frame deletion of rcc02111 without its first 4 and last 4 codons.
Gmr This work
aR.capsulatus strain MT1131 is derived from SB1003 in multiple steps, as described in Zannoni et al. (1980): first a Ps-deficient mutant (TL1) was obtained using tetracycline suicide, then its crtD derivative was constructed by GTA cross to yield MT113, and then its Ps-proficient derivative was obtained via a second GTA cross.
S- 8
Table S2. Primers used in this work.
Gm-F 5-AAGCTTGTCGACCCGATCTGAGC-5
Gm-R 5- TCTAGAACTAGTGGATCCCCCG-3
01423-F1 5-CGGCCAGTGAATTCGAGCTCGGTACATGCGCAGGGTATAGGTGACGCC-3
01423-R1 5-TCATTTTTCGAACTGCGGGTGGCTCCAGGCGTTTTCTTCTTCGAAGTGCC-3
01423-F2 5-GAGCCACCCGCAGTTCGAAAAATGATCCGACTTCGCAAGCCAGGTCC-3
01423-R2 5-AAGCTTGCATGCCTGCAGGTCGACTGCGATCGTCTTTGCCGATCTGGC-3
01423Gm-R1 5-GAAAATGGCTCAGATCGGGTCGACAAGCTTAAGTGGCAGACATGACATGTCTCC-3
01423Gm-F3 5-TTCCCGGCCGGGGGATCCACTAGTTCTAGAGAAGAAAACGCCTGATCCGACTTCG -3
01423Su-F1 5-TACGCCAAGCTTGCATGCCTGCAGGGTCTGTTGCGCCAGCACGGCTTTGC-3
01423Su-R1 5-AGTGGCAGACATGACATGTCTCCTC-3
01423Su-F2 5-GAGGAGACATGTCATGTCTGCCACTGAAGAAAACGCCTGATCCGACTTCG-3
01423Su-R2 5-AAACGACGGCCAGTGAATTCGAGCTGGCGACGGACAATTGCCGGGGCTG-3
00738-F1 5-CGGCCAGTGAATTCGAGCTCGGTACACTGCGCCAAGACCTGCCCCAAGG-3
00738-R1 5-AAAAATGGCTCAGATCGGGTCGACAAGCTTAGACGATGCGGAAGAAGCAGCCCG -3
00738-F2 5-TTCCCGGCCGGGGGATCCACTAGTTCTAGAGATCTGGGCGGCGCGCTCTCGGC-3 00738-R2 5-AAGCTTGCATGCCTGCAGGTCGACTTGACCATGCTGCTGGAAGACGAC-3 00891-F1 5-CGGCCAGTGAATTCGAGCTCGGTACAGATGGTGATCCGCGCCATCGCG-3
00891-R1 5-AAAAATGGCTCAGATCGGGTCGACAAGCTTCGCCGAGGCCGAGAGATGTTCGG -3
00885-F2 5-TTCCCGGCCGGGGGATCCACTAGTTCTAGAACCCGAACCATCACTGCCCCTTCTG-3
00885-R2 5-AAGCTTGCATGCCTGCAGGTCGACTACGCCGCCGGATGGCATTCAAAAGG-3
01027-F1 5-CGGCCAGTGAATTCGAGCTCGGTACCCGCCCGAAAGCGCCAGAAGCGC-3
01027-R1 5-AAAAATGGCTCAGATCGGGTCGACAAGCTTGCTGCCCGTAGGCGTCGGCAACC-3
01031-F2 5-TTCCCGGCCGGGGGATCCACTAGTTCTAGAGCCGACCGGGTTCTGGTGCTGGC-3
01031-R2 5-AAGCTTGCATGCCTGCAGGTCGACTGCGATCGCGCAATTCGGCCTTGGC-3 02111Su-F1 5-TACGCCAAGCTTGCATGCCTGCAGGATCGAGCTGTGGCAGGTGATGCGC-3 02111Su-R1 5-CAGAATCCGCATGGGAGGAGCATGGC-3
02111Su-F2 5-CCATGCTCCTCCCATGCGGATTCTGCCGCCCCGCCTCTGATCTTCGCC-3
02111Su-R2 5-AAACGACGGCCAGTGAATTCGAGCTGGCAGCGGCACCACCGGGTTGCC-3
02109-R1 5-AAAAATGGCTCAGATCGGGTCGACAAGCTTAAGGCACCCGGCGACAAGGGCC-3
S- 9
02109-F2 5-TTCCCGGCCGGGGGATCCACTAGTTCTAGACTGATCACGGCGGACGGGCAAACC-3
02109-R2 5-AAGCTTGCATGCCTGCAGGTCGACTGTTCTAACCTTAGGCCGCCAGAC-3
00098Su-F1 5-TACGCCAAGCTTGCATGCCTGCAGGCGACGCTGGGCCTTGACCCGGAG-3
00098Su-R1 5-AAGGCGGCGCATGGGTGTCTCCAG-3
00098Su-F2 5-ACTGGAGACACCCATGCGCCGCCTTATCCGTTTCTGAGATCCGTTTCC-3
00098Su-R2 5-AAACGACGGCCAGTGAATTCGAGCTTCGTCCATGCGCAGATGCAGATC-3
01445-F1 5-CGGCCAGTGAATTCGAGCTCGGTACCGCTGGTGGCAGAACCGATCAGC-3
01445-R1 5-AAAAATGGCTCAGATCGGGTCGACAAGCTTGGTGATGACGATCGTGCCCAGCG-3
01445-F2 5-TTCCCGGCCGGGGGATCCACTAGTTCTAGAGACGGCGCTTTTGATCGACGCGC-3
01445-R2 5-AAGCTTGCATGCCTGCAGGTCGACTTGGCATGGATGCGATGCTGATCG-3
03065-F1 5-CGGCCAGTGAATTCGAGCTCGGTACTCCAGCGGCAGATGCGTGGCGGC-5
03065-R1 5-AAAAATGGCTCAGATCGGGTCGACAAGCTTCGGCGGCGCGCATCCAGAACACC-3
03067-F2 5-TTCCCGGCCGGGGGATCCACTAGTTCTAGATTCGGCCCGATGGTCGAGGACCGC-3
03067-R2 5-AAGCTTGCATGCCTGCAGGTCGACtCCAGCCGATGCGCGATGACATTCG-3
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Table S3. Excel spreadsheet compiled datasets. Data related to the identification, annotations,
quantitation and statistics of R. capsulatus proteins are cumulated in Table S3. Sheet 1 (R.
capsulatus Proteins): Full genome listing of R. capsulatus (3632 entries, UniprotKB). Various
annotations are included: PsortB localizations, TIGRFAM and TIGR roles, GO annotations,
Interpro, and KEGG classifications, as well as protein identifiers. Those proteins experimentally
identified in the current study are marked with asterisks (first column), and those unidentified are
designated in gray colored rows. Sheet 2 (Identified Proteins): Proteins experimentally identified
in the current study (2430 entries), with the number of unique peptides, MS/MS counts and sequence
coverage listed along with other information. Sheet 3 (Unidentified Proteins): Proteins unidentified
in the present study are listed along with functional annotations where available. Sheet 4 (Single
Peptide-ID): List of proteins identified with only one unique peptide (59 entries). Those proteins
identified with single MS/MS spectra are highlighted in yellow (13 entries). Annotated spectra for
all single peptides are provided in Figure S4. Sheet 5 (Quantified-Statistics): The 1926 proteins
quantified in this study are listed, along with T-test p-values and Fold-changes for +Cu vs control
and for +BCS vs control (see Experimental Procedures). The 75 proteins that were significantly
affected by changes in Cu levels (i.e., surpassed the criteria of p<0.05 and FC>2.0) are highlighted
as noted in the table headings.
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Supplemental FIGURES
Figure S1. Nadi staining of R. capsulatus colonies grown under different Cu concentrations. Cu-sufficient (control), Cu-excess
(+Cu) and Cu-depleted (+BCS) refer to MedA medium containing 0.3 µM Cu, 5 µM Cu and 5mM BCS, respectively.
Figure S1. Selamoglu et al.
Cu-sufficient(control)
Cu-excess(+Cu)
Cu-depleted(+BCS)
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Figure S2. Pearson correlation for individual experimental runs. Color-coded Pearson r-values are shown for the 31 individual runs
using lysis/digestion protocol A and Q-Exactive mass spectrometer (Experimental Procedures). The samples are identified by Cu
condition (Ctrl, +Cu and +BCS), biological repeat (B1, B2, B3) and technical repeat (R1, R2, R3, R4). All r values were consistently
high, within 0.92 to 0.99, except the sample +Cu(B2R1) that had r-values between 0.75-0.83 and was subsequently discarded.
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Figure S3. The NEBuilder HiFi DNA Assembly Method. (a) the version used for GTA mediated chromosomal inactivation, and (b)
that used for construction of chromosomal in-frame seamless deletions of R. capsulatus chromosomal genes. The dotted lines indicate
the overlapping regions between the vector arms and the DNA fragments being inserted.
a)Vector: pRK415 (KpnI/XbaI), 10.5 kb
GmR cassette20-30 nt
overlapping sequence
N-terminal part of gene
C-terminal part of gene
-Kpn
I
-Xba
I
20-30 ntoverlapping sequence
5’
-3’
3’3’
5’
5’ 5’
3’
3’
5’
5’
5’3’
3’3’ 5’
TetR
b)Vector: pZDJ (SalI/SacI), 4.8 kb
25 ntoverlapping sequence
Upstream of gene-(5’ flanking region)
-Sal
I
-Sac
I
ATG NNN NNN NNN
ATG NNN NNN NNN NNN NNN NNN TGA
TAC NNN NNN NNN
TAC NNN NNN NNN NNN NNN NNN ACT
Downstream of gene(3’ flanking region)
Start and first 3 codons
Last 3 codons and Stop 25 ntoverlapping sequence
-3’
3’
3’ 5’
5’
5’3’ 5’
-5’
5’
3’
3’
GmRsacB
Figure S3. Selamoglu et al.
S- 14
Figure S4. Annotated spectra corresponding to proteins identified by a single unique peptide (see Sheet 4 (Single Peptide-ID) in Table S3).
y₁-NH₃130.0863
y₁147.1128
b₂-H₂O199.0713
y₂-NH₃258.1448
y₂275.1714
y₃346.2085
b₄-H₂O357.1405
y₄417.2456
b₅-H₂O486.1831
y₅504.2776
b₆-H₂O557.2202
y₆575.3148
y₇646.3519
y₈717.389
y₉846.4316
y₁₀917.4687
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
1.61.8
2
[1e4]
200 400 600 800
Raw File Scan Method Score m/zMT1131-minusCu-Set3-02 10728 FTMS; HCD 56.23 710.83
- E S G T E A
y₁₀
E
y₉
A
y₈
A
y₇
A
y₆
S
y₅
A
y₄
A
y₃
Q
y₂
K
y₁
-
a₂143.1179
b₂171.1128
y₁175.119
b₃299.1714
b₄430.2119
b₅501.249
b₆-NH₃583.2908
b₆600.3174
y₆798.4621
y₇-NH₃878.4883
y₇895.5148
y₈1008.599
y₉1107.667
y₁₀1178.704
y₁₁1309.745
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
1.6
[1e5]
200 400 600 800 1000 1200
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-Set3-01 42715 FTMS; HCD 59.07 804.76
- I Gb₂
Qb₃
M
y₁₁
b₄A
y₁₀
b₅V
y₉
b₆L
y₈
P
y₇
P
y₆
W L N L R
y₁
-
y₁-NH₃158.0924
b₂-NH₃169.0608
y₁175.119
b₂186.0873
y₂-NH₃257.1608
b₃-NH₃266.1135
y₂274.1874
b₃283.1401
b₄-NH₃379.1976
y₃-NH₃385.2194
b₄396.2241
y₃402.2459
y₄-H₂O471.2674
y₄-NH₃472.2514
y₄489.278
b₅-NH₃507.2562
b₅524.2827
y₅-H₂O584.3515
y₅-NH₃585.3355
y₅602.362
b₆-NH₃654.3246
b₆671.3511
y₆-H₂O731.4199
y₆-NH₃732.4039
y₆749.4304
y₇-H₂O859.4785
y₇-NH₃860.4625
y₇877.489
y₈-NH₃973.5465
y₈990.5731
y₉-NH₃1070.599
y₉1087.626
020
4060
80100
120
00.5
11.5
22.5
33.5
[1e6]
200 400 600 800 1000 1200
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-Set2-02 33683 FTMS; HCD 215.14 636.86
- A Nb₂
P
y₉
b₃I
y₈
b₄Q
y₇
b₅F
y₆
b₆L
y₅
S
y₄
Q
y₃
V
y₂
R
y₁
-
y₁-NH₃158.0924
a₂161.0743
y₁175.119
b₂189.0692
b₃288.1376
y₂289.1619
b₄-NH₃385.154
b₄402.1806
y₃402.2459
b₅-NH₃484.2224
b₅501.249
y₄533.2864
b₆-H₂O584.2861
b₆-NH₃585.2701
b₆602.2967
y₅-NH₃644.3185
y₅661.345
b₁₃²⁺685.3156
y₆-NH₃715.3556
y₆732.3821
b₈-NH₃785.3498
y₇-NH₃862.424
y₇879.4505
b₉-H₂O915.4063
b₁₈²⁺949.4428
y₈-NH₃977.4509
y₈994.4775
y₉-NH₃1048.488
y₉1065.515
b₁₀1070.476
y₁₉²⁺1088.02
y₁₀-H₂O1175.563
y₁₀-NH₃1176.547
y₁₀1193.573
b₁₁1199.518
b₁₂1256.54
y₁₁1294.621
b₁₃1369.624
y₁₂1407.705
y₁₄1577.81
y₁₆1843.912
y₁₈2045.99
y₂₀-NH₃2259.053
y₂₀2276.08
y₂₁2375.148
y₂₂2489.191
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
1.61.8
[1e5]
500 1000 1500 2000 2500
Raw File Scan Method Score m/zMT1131-plusCu-Set2-01 41976 FTMS; HCD 148.95 926.45
- M Gb₂
Vb₃
N
y₂₂
b₄V
y₂₁
b₅T
y₂₀
b₆E
y₁₉²⁺
A
y₁₈
M H
y₁₆
b₁₀Eb₁₁
G
y₁₄
b₁₂L
b₁₃L
y₁₂
T
y₁₁
Q
y₁₀
A
y₉
D
y₈
b₁₈²⁺
F
y₇
A
y₆
Q
y₅
M
y₄
I
y₃
N
y₂
R
y₁
-
y₄-NH₃411.24
y₄428.26
b₆525.27
y₅-NH₃597.31
y₅614.34
b₇624.34
y₆727.42
b₈752.39
y₇798.46
b₉809.42
y₈869.5
b₁₀922.5
y₉982.58
y₁₀1111.63
b₁₂1122.58
y₁₁-H₂O1164.65
y₁₁1182.66
b₁₃1235.66
b₁₄1306.7
y₁₃1352.77
b₁₅1377.74
y₁₄1480.83
b₁₆1490.82
y₁₅1579.9
b₁₇-H₂O1658.89
b₁₇-NH₃1659.87
b₁₇1676.9
y₁₆1678.96
b₁₈-H₂O1755.94
y₁₇1793.99
b₁₉-NH₃1813.95
b₁₉1830.97
b₂₀1930.04
y₁₉-NH₃1945.05
020
4060
80100
120
0200
400600
8001000
12001400
400 600 800 1000 1200 1400 1600 1800 2000
Raw File Scan Method Score m/zCTRL-minusCu-Biorep1-Run2 23785 ITMS; CID 154.46 1053.09
- A A P A D
y₁₇
V
y₁₆
b₆V
y₁₅
b₇Q
y₁₄
b₈G
y₁₃
b₉I
b₁₀A
y₁₁
E
y₁₀
b₁₂I
y₉
b₁₃A
y₈
b₁₄A
y₇
b₁₅L
y₆
b₁₆W
y₅
b₁₇P
y₄
Gb₁₉
Vb₂₀
R -
y₁-NH₃158.09
y₁175.12
a₂187.11
b₂-H₂O197.09
y₂-NH₃215.11
y₂232.14
b₃286.14
y₄-NH₃385.22
y₄402.25
b₄417.18
y₅-NH₃498.3
y₅515.33
b₅530.26
y₆628.41
b₆643.35
y₇-NH₃742.43
y₈-NH₃813.47
y₈830.49
b₉870.48
020
4060
80100
120
01
23
45
6
[1e5]
200 400 600 800 1000
Raw File Scan Method Score m/zPlusBC-Biorep1-Run1 10767 ITMS; CID 114.5 523.3
- D Va₂
A
y₈
b₃Mb₄
L
y₆
b₅L
y₅
b₆G
y₄
L G
y₂
b₉R
y₁
-
y₁132.1019
a₂200.1394
b₂-NH₃211.1077
y₂-H₂O215.139
b₂228.1343
y₂233.1496
b₃-NH₃308.1605
b₄-NH₃365.1819
y₄-H₂O425.2758
y₄443.2864
y₅-H₂O553.3708
y₅571.3814
y₆-H₂O682.4134
y₆700.424
b₇-NH₃719.3723
y₇-H₂O739.4349
y₇757.4454
y₈-H₂O836.4876
y₈854.4982
020
4060
80100
120
00.5
11.5
22.5
[1e5]
100 200 300 400 500 600 700 800 900
Raw File Scan Method Score m/zMT1131-minusCu-Set3-02 21643 FTMS; HCD 83.31 541.32
- L Nb₂
P
y₈
G
y₇
E
y₆
K
y₅
P
y₄
I T
y₂
L
y₁
-
y₁-NH₃158.0924
a₂169.1335
y₁175.119
b₂197.1285
y₂246.1561
b₃268.1656
b₄325.187
y₃333.1881
y₄-NH₃430.2045
y₄447.231
b₅456.2275
y₅-NH₃501.2416
y₅518.2681
b₆-H₂O539.2646
b₆557.2752
y₆631.3522
y₇702.3893
b₈-H₂O737.3651
y₈865.4526
b₁₀883.4342
y₉922.4741
y₁₀-NH₃1006.495
y₁₀1023.522 y₁₁
1094.559
y₁₂1151.58
b₁₃1204.567
y₁₃-H₂O1234.617
y₁₃1252.628
y₁₄-H₂O1331.67
y₁₄1349.681
y₁₅-H₂O1432.718
y₁₅-NH₃1433.702
y₁₅1450.728
y₁₆1581.769
y₁₇1638.79
y₁₈1709.828
y₁₉-H₂O1788.87
y₁₉1806.88
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
1.61.8
[1e6]
200 400 600 800 1000 1200 1400 1600 1800
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-01 26292 FTMS; HCD 150.12 953.98
- V P
y₁₉
b₂A
y₁₈
b₃G
y₁₇
b₄M
y₁₆
b₅T
y₁₅
b₆P
y₁₄
T
y₁₃
G
y₁₂
A
y₁₁
b₁₀T
y₁₀
G
y₉
Y
y₈
b₁₃A
y₇
L
y₆
A
y₅
N
y₄
S
y₃
A
y₂
R
y₁
-
y₁-NH₃130.0863
y₁147.1128
y₂-NH₃187.1077
a₂203.1026
y₂204.1343
b₂-H₂O213.087
b₂231.0975
y₃305.1819
b₃-H₂O310.1397
b₃328.1503
b₄-H₂O409.2082
y₄-H₂O415.23
y₄-NH₃416.214
b₄427.2187
y₄433.2405
b₅-H₂O480.2453
b₅498.2558
y₅532.3089
y₆-H₂O611.3511
y₆629.3617
b₇694.377
y₇-H₂O740.3937
y₇758.4043
b₈795.4247
y₈857.4727
y₉-H₂O940.5098
y₉-NH₃941.4938
y₉958.5204
y₁₀1057.589
y₁₁-H₂O1136.631
y₁₁-NH₃1137.615
y₁₁1154.642
y₁₂-H₂O1207.668
y₁₂1225.679
y₁₃1324.747
y₁₄1421.8
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
[1e6]
200 400 600 800 1000 1200 1400 1600
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-Set2-01 22555 FTMS; HCD 175.48 826.45
- T Eb₂
P
y₁₄
b₃V
y₁₃
b₄A
y₁₂
b₅P
y₁₁
V
y₁₀
b₇T
y₉
b₈V
y₈
E
y₇
P
y₆
V
y₅
Q
y₄
T
y₃
G
y₂
K
y₁
-
y₃439.16
b₆441.25
y₄496.19
b₇512.28
b₁₃²⁺526.79
y₁₀²⁺534.72
y₅553.21
b₉640.34
y₆666.29
y₁₄²⁺677.78
y₇723.31
b₁₀753.43
b₁₈²⁺776.39
b₁₉²⁺849.91
b₁₁866.51 b₂₁²⁺
934.96
b₁₂937.55
y₁₀-NH₃1051.4
y₁₀1068.42
b₂₅²⁺1091.05
b₂₇²⁺1155.08
y₁₁1169.47
b₁₅-H₂O1176.64
b₁₅1194.65
b₂₉²⁺1247.14
y₁₃1297.53
b₁₆1309.67
y₁₄-NH₃1337.52
b₃₂²⁺1339.68
y₁₄1354.55
b₁₇1437.73
y₃₁²⁺1455.64
y₁₆1538.67
b₃₆²⁺1562.76
y₁₇-H₂O1591.7
y₃₄²⁺1604.25
y₁₈-NH₃1649.7
y₁₈1666.73
y₄₀²⁺1796.33
b₂₀1797.88
y₂₀1850.85
b₂₁-NH₃1851.89
b₂₁1868.92
b₂₃1996.98
020
4060
80100
120
050
100150
200250
300
400 600 800 1000 1200 1400 1600 1800 2000
Raw File Scan Method Score m/zPlusBC-Biorep2-Run1 34218 ITMS; CID 74.87 962.96
- A L A G
y₄₀²⁺
G Ab₆
Ab₇
G Ab₉
I
y₃₄²⁺
b₁₀I
b₁₁Ab₁₂
D
y₃₁²⁺
b₁₃²⁺A A
b₁₅Db₁₆
Qb₁₇
Nb₁₈²⁺
Mox
b₁₉²⁺Vb₂₀
Ab₂₁
G Ab₂₃
A
y₂₀
Lb₂₅²⁺
G
y₁₈
Ab₂₇²⁺
L
y₁₆
Ab₂₉²⁺
G
y₁₄
G
y₁₃
Ab₃₂²⁺
T
y₁₁
C
y₁₀
N Ab₃₆²⁺
G
y₇
I
y₆
G
y₅
G
y₄
P
y₃
C Y -
a₂145.0972
b₂-H₂O155.0815
y₁-NH₃158.0924
b₂173.0921
y₁175.119
b₃-H₂O226.1186
b₃244.1292
b₄-H₂O297.1557
y₃417.2456
y₆676.326
y₇773.3788
y₈-NH₃853.405
y₈870.4316
y₉941.4687
y₁₀1012.506
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
[1e5]
200 400 600 800 1000 1200 1400
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-01 9780 FTMS; HCD 60.1 592.32
- A Tb₂
A
y₁₀
b₃A
y₉
P
y₈
P
y₇
S
y₆
G D I
y₃
E R
y₁
-
y₁-NH₃158.0924
y₁175.119
a₂201.1234
b₂229.1183
y₂-NH₃286.151
b₃300.1554
y₂303.1775
b₄-H₂O369.1769
y₄417.2205
y₅504.2525
y₆617.3365
y₇704.3686
y₈-H₂O757.3951
y₈775.4057
020
4060
80100
120
01
23
45
6
[1e4]
100 200 300 400 500 600 700 800
Raw File Scan Method Score m/zMT1131-plusCu-Set3-02 14106 FTMS; HCD 66.69 502.26
- D Ib₂
A
y₈
b₃S
y₇
L
y₆
S
y₅
G
y₄
G Q
y₂
R
y₁
-
y₁-NH₃158.0924
y₁175.119
b₂-NH₃253.1659
b₂270.1925
y₂288.203
y₃359.2401
b₄424.2667
y₄430.2772
y₅531.3249
b₅553.3093
y₁₁²⁺609.304
b₆650.362
y₆660.3675
b₇721.3991
y₇761.4152
y₈-H₂O872.4472
y₈890.4578
b₉-H₂O929.4839
b₉947.4945
y₉-H₂O973.4949
y₉991.5055
b₁₀-NH₃1031.516
y₁₀-H₂O1102.537
y₁₀-NH₃1103.522
y₁₀1120.548
b₁₁1177.585
y₁₁-H₂O1199.59
y₁₁1217.601
020
4060
80100
120
00.2
0.40.6
0.81
[1e5]
200 400 600 800 1000 1200 1400 1600 1800
Raw File Scan Method Score m/zMT1131-minusCu-Set2-01 18462 FTMS; HCD 96.14 647
- L Rb₂
P Gb₄
Eb₅
Pb₆
Ab₇
P
y₁₁
E
y₁₀
b₉T
y₉
E
y₈
b₁₁T
y₇
E
y₆
T
y₅
A
y₄
A
y₃
L
y₂
R
y₁
-
b₂115.0502
y₁-NH₃158.0924
y₁175.119
b₃229.0931
y₂288.203
b₄342.1772
y₃389.2507
b₅-NH₃396.1878
b₅413.2143
b₆-NH₃524.2463
b₆541.2729
y₄552.314
b₇-NH₃637.3304
y₅653.3617
b₈-H₂O751.3733
y₇-H₂O820.4312
y₇838.4417
y₈953.4687
y₉1066.553
y₁₀1181.58
y₁₁1294.664
y₁₂-NH₃1405.696
y₁₂1422.722
y₁₃1493.759
020
4060
80100
120
01
23
45
67
89
[1e4]
200 400 600 800 1000 1200 1400 1600 1800
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-Set3-02 31927 FTMS; HCD 99.01 917.97
- G Gb₂
Nb₃
Lb₄
A
y₁₃
b₅Q
y₁₂
b₆I
y₁₁
D
y₁₀
L
y₉
D
y₈
G
y₇
Q T
y₅
Y
y₄
T
y₃
L
y₂
R
y₁
-
y₁-NH₃158.0924
y₁175.119
a₂201.1234
b₂-H₂O211.1077
b₂229.1183
b₃-H₂O339.1663
b₃357.1769
y₄-NH₃373.183
y₄390.2096
b₄-H₂O452.2504
b₄470.2609
y₅503.2936
y₆-H₂O582.3358
y₆-NH₃583.3198
y₆600.3464
y₇-H₂O681.4042
y₇699.4148
y₈812.4989
y₉-H₂O922.5469
y₉940.5574
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
1.61.8
[1e5]
200 400 600 800 1000 1200
Raw File Scan Method Score m/zMT1131-minusCu-Set2-01 25381 FTMS; HCD 111.61 585.34
- E Vb₂
Q
y₉
b₃L
y₈
b₄V
y₇
P
y₆
L
y₅
S
y₄
G A R
y₁
-
y₁175.119
b₂187.1077
b₃-H₂O270.1448
y₂303.1775
y₃440.2364
y₄527.2685
y₅640.3525
y₆-H₂O723.3896
y₆741.4002
y₇-NH₃852.4322
y₇869.4588
y₈984.4857
y₉1055.523
020
4060
80100
120
00.5
11.5
22.5
[1e4]
200 400 600 800 1000 1200
Raw File Scan Method Score m/zMT1131-plusCu-Set3-01 6420 FTMS; HCD 79.97 671.84
- V Sb₂
T A
y₉
D
y₈
Q
y₇
T
y₆
L
y₅
S
y₄
H
y₃
Q
y₂
R
y₁
-
y₁-NH₃158.0924
a₂159.0764
y₁175.119
b₂187.0713
y₂312.1779
b₃316.1139
b₄387.151 y₃
425.2619
y₄554.3045
y₅668.3474
y₆-H₂O778.3955
y₆-NH₃779.3795
y₆796.406
y₇-NH₃907.4381
y₇924.4646
y₈995.5017
y₉1066.539
020
4060
80100
120
00.5
11.5
22.5
3
[1e4]
200 400 600 800 1000
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-Set2-01 7984 FTMS; HCD 74.84 691.33
- D Ab₂
Eb₃
A
y₉
b₄A
y₈
Q
y₇
Q
y₆
N
y₅
E
y₄
L
y₃
H
y₂
R
y₁
-
a₂157.1335
y₁-NH₃158.0924
y₁175.119
b₂185.1285
b₃282.1812
y₆²⁺379.6826
b₄395.2653
y₄534.2671
y₉²⁺549.7882
y₅-H₂O603.2885
y₁₀²⁺606.3302
y₅621.2991
y₆-H₂O740.3474
y₆758.358
y₁₅²⁺867.4885
y₇-H₂O868.406
y₇-NH₃869.39
y₇886.4166
y₈999.5007
y₉-NH₃1081.543
y₉1098.569
y₁₀-NH₃1194.627
y₁₀1211.653
y₁₁1339.712
y₁₂-NH₃1419.738
y₁₂1436.764
y₁₃-H₂O1489.791
y₁₃1507.802
y₁₄1620.886
020
4060
80100
120
00.2
0.40.6
0.81
1.2
[1e6]
200 400 600 800 1000 1200 1400 1600
Raw File Scan Method Score m/zMT1131-plusCu-01_150512102711 44969 FTMS; HCD 73.17 672.72
- I Ab₂
Pb₃
L
y₁₅²⁺
b₄L
y₁₄
A
y₁₃
P
y₁₂
Q
y₁₁
I
y₁₀
V
y₉
I
y₈
Q
y₇
H
y₆
S
y₅
P
y₄
F D R
y₁
-
y₁-NH₃158.0924
y₁175.119
a₂187.1441
b₂-H₂O197.1285
b₂215.139
y₂-NH₃229.1295
y₂246.1561 y₃-NH₃
392.1928
y₃409.2194
y₄480.2565
y₆608.3151
y₇-NH₃648.31
y₇665.3365
y₈-NH₃776.3686
y₈793.3951
y₉894.4428
020
4060
80100
120
01
23
45
[1e4]
200 400 600 800
Raw File Scan Method Score m/zMT1131-minusCu-01 6607 FTMS; HCD 84.57 504.27
- L T
y₉
b₂Q
y₈
G
y₇
G
y₆
A A
y₄
Y
y₃
A
y₂
R
y₁
-
y₂246.16
b₃258.14
y₃361.18
y₄-H₂O458.2
y₄476.21
b₅-H₂O532.24
b₅550.25
y₅623.28
y₆-H₂O676.3
y₆-NH₃677.29
b₆-H₂O692.27
y₆694.32
b₆710.28 y₇-NH₃
791.33
y₇808.36
b₈-H₂O896.36
b₈914.37
y₈955.43
b₉-NH₃1011.39
y₉1012.45
b₉1028.41
b₁₀-H₂O1081.44
b₁₀-NH₃1082.42
b₁₀1099.45
y₁₀-NH₃1155.45
y₁₀1172.48
b₁₁-NH₃1229.49
b₁₁1246.52
y₁₁-H₂O1283.51
y₁₁-NH₃1284.49
y₁₁1301.52
b₁₂1361.55
y₁₂1464.58
b₁₃1476.57
b₁₄-NH₃1530.58
y₁₃1535.62
b₁₄1547.61
y₁₄-NH₃1605.63
y₁₄1622.65
020
4060
80100
120
01
23
45
[1e4]
200 400 600 800 1000 1200 1400 1600
Raw File Scan Method Score m/zPlusCu-Biorep2-Run1 13938 ITMS; CID 220.37 861.87
- V S
y₁₄
A
y₁₃
b₃Y
y₁₂
E
y₁₁
b₅C
y₁₀
b₆G
y₉
F
y₈
b₈N
y₇
b₉A
y₆
b₁₀F
y₅
b₁₁D
y₄
b₁₂D
y₃
b₁₃A
y₂
b₁₄R -
y₁175.119
b₄-H₂O271.1037
y₂272.1717
y₅553.3457
y₆640.3777
y₇753.4618
y₉1029.609
y₁₁1199.715
y₁₂1314.742
y₁₃1385.779
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
1.6
[1e4]
200 400 600 800 1000 1200 1400
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-Set3-03 33767 FTMS; HCD 51.21 958.52
- S G G S L Q A
y₁₃
D
y₁₂
A
y₁₁
V Y
y₉
L I
y₇
S
y₆
P
y₅
A L P
y₂
R
y₁
-
y₁-NH₃158.0924
a₂173.1285
y₁175.119
b₂-H₂O183.1128
b₂201.1234
y₂288.203
b₃348.1918
y₃389.2507
b₄-H₂O445.2082
b₄463.2187
y₄552.314
b₅560.2715
y₅665.3981
b₆-H₂O671.3035
b₆689.3141
y₆778.4822
b₇-H₂O784.3876
b₇802.3981
y₇835.5036
b₈899.4509
y₈948.5877
y₉1063.615
b₁₀1112.562
b₁₁-NH₃1208.62
y₁₀1210.683
y₁₁1323.767
y₁₂1437.81
y₁₃1536.878
y₁₄-NH₃1616.905
y₁₄1633.931
y₁₅1747.015
y₁₇1973.111
020
4060
80100
120
00.5
11.5
22.5
[1e6]
500 1000 1500 2000 2500
Raw File Scan Method Score m/zMT1131-plusCu-01_150512102711 67532 FTMS; HCD 124.92 1218.67
- T Vb₂
Fb₃
Db₄
P
y₁₇
b₅Eb₆
I
y₁₅
b₇P
y₁₄
b₈V
y₁₃
N
y₁₂
b₁₀I
y₁₁
F
y₁₀
D
y₉
L
y₈
G
y₇
L
y₆
I
y₅
Y
y₄
T
y₃
I
y₂
R
y₁
-
y₁-NH₃158.0924
a₂173.0921
y₁175.119
b₂-H₂O183.0764
b₂201.087
y₂-NH₃257.1608
b₃-H₂O298.1034
y₃-NH₃314.1823
b₃316.1139
y₃331.2088
b₄-H₂O369.1405
y₄-NH₃443.2249
y₄460.2514
y₅531.2885
y₆-H₂O600.31
y₆618.3206
y₇-H₂O731.3505
y₇749.361
y₈820.3982
y₉935.4251
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
[1e5]
200 400 600 800
Raw File Scan Method Score m/zMT1131-minusCu-Set2-02 11862 FTMS; HCD 103.55 568.26
- E Ab₂
D
y₉
b₃A
y₈
M
y₇
S
y₆
A
y₅
E
y₄
G
y₃
V R
y₁
-
y₁-NH₃158.0924
y₁175.119
a₂189.087
b₂217.0819
b₃-H₂O296.1241
y₂303.2139
b₃314.1347
b₄-H₂O367.1612
b₄385.1718
y₃416.298
y₄529.382
y₅616.4141
b₆-H₂O627.2773
b₆645.2879
b₇-H₂O728.325
y₆729.4981
b₇746.3355
y₇844.5251
y₈915.5622
y₉1002.594
y₁₀-H₂O1085.631
y₁₀-NH₃1086.615
y₁₀1103.642
y₁₁-H₂O1248.695
y₁₁-NH₃1249.679
y₁₁1266.705
y₁₂-H₂O1345.747
y₁₂1363.758
y₁₃1434.795
y₁₄-H₂O1513.837
y₁₄1531.848
y₁₅-H₂O1642.88
020
4060
80100
120
01
23
45
67
8
[1e6]
200 400 600 800 1000 1200 1400 1600
Raw File Scan Method Score m/zMT1131-plusCu-03 26579 FTMS; HCD 137.6 874.47
- S Eb₂
P
y₁₄
b₃A
y₁₃
b₄P
y₁₂
Y
y₁₁
b₆T
y₁₀
b₇S
y₉
A
y₈
D
y₇
L
y₆
S
y₅
L
y₄
I
y₃
K
y₂
R
y₁
-
y₁-NH₃130.0863
y₁147.1128
a₂173.1285
b₂-H₂O183.1128
b₂201.1234
y₂-H₂O230.1499
y₂248.1605
b₃-H₂O312.1554
b₃330.166
y₃-H₂O359.1925
y₃377.2031
y₄-H₂O416.214
y₄434.2245
b₄-H₂O441.198
b₄459.2086
b₅-H₂O512.2351
y₅-H₂O529.298
b₅530.2457
y₅547.3086
y₆-H₂O644.325
y₆-NH₃645.309
y₆662.3355
y₇-H₂O807.3883
y₇825.3989
y₈-H₂O878.4254
y₈896.436
y₉967.4731
y₁₀-H₂O1078.505
y₁₀1096.516
y₁₁-H₂O1207.548
y₁₁1225.558
020
4060
80100
120
00.5
11.5
22.5
33.5
4
[1e5]
200 400 600 800 1000 1200 1400 1600
Raw File Scan Method Score m/zMT1131-minusCu-Set3-02 25432 FTMS; HCD 188.49 713.34
- S Lb₂
E
y₁₁
b₃E
y₁₀
b₄A
y₉
b₅A
y₈
Y
y₇
D
y₆
L
y₅
G
y₄
E
y₃
T
y₂
K
y₁
-
a₂175.09
b₂203.08
b₃274.12
b₄345.16
b₅416.2
y₃417.25
y₄516.31
y₅-NH₃600.34
b₇601.28
b₈-NH₃685.3
y₆745.42
y₇-H₂O784.43
y₇-NH₃785.42
y₇802.44
y₈-H₂O855.47
y₈-NH₃856.45
y₈873.48
b₁₁-H₂O1025.51
b₁₁-NH₃1026.49
b₁₁1043.52
020
4060
80100
120
01000
20003000
40005000
60007000
80009000
200 400 600 800 1000 1200
Raw File Scan Method Score m/zCTRL-minusCu-Biorep1-Run3 4792 ITMS; CID 83.65 609.82
- A Mb₂
Ab₃
Ab₄
A
y₈
b₅G
y₇
Q
y₆
b₇T V
y₄
E
y₃
Ib₁₁
R -
y₁-NH₃158.0924
a₂173.1285
y₁175.119
b₂-H₂O183.1128
b₂201.1234
y₂-NH₃287.135
y₂304.1615
y₃-H₂O415.1936
y₃433.2041
y₄504.2413
y₅591.2733
y₆-H₂O701.3213
y₆-NH₃702.3053
y₆719.3319 y₇
820.3795
y₈-NH₃890.385
y₈907.4116
020
4060
80100
120
01
23
45
[1e4]
200 400 600 800 1000
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-Set2-02 5857 FTMS; HCD 78.15 510.75
- L S
y₈
b₂T
y₇
Q
y₆
S
y₅
A
y₄
E
y₃
E
y₂
R
y₁
-
y₁-NH₃158.0924
a₂173.0921
y₁175.119
b₂201.087
b₃330.1296
y₂338.1823
b₄443.2136
y₄-NH₃547.2511
y₄564.2776
b₆585.2879
y₅677.3617
y₆748.3988
y₇819.4359
y₈-NH₃873.4465
y₈890.473
y₉1003.557
y₁₀1132.6
y₁₁-NH₃1244.616
020
4060
80100
120
00.2
0.40.6
0.81
[1e5]
200 400 600 800 1000 1200 1400 1600
Raw File Scan Method Score m/zMT1131-plusCu-02 16394 FTMS; HCD 63.69 666.84
- A Eb₂
E
y₁₀
b₃L
y₉
b₄A
y₈
A
y₇
b₆A
y₆
L
y₅
P
y₄
E Y
y₂
R
y₁
-
y₁-NH₃158.0924
y₁175.119
a₂187.1441
b₂-H₂O197.1285
b₂215.139
y₂-NH₃286.151
b₃-H₂O298.1761
y₂303.1775
b₃316.1867
b₄-H₂O399.2238
y₃-H₂O414.2096
y₃-NH₃415.1936
y₃432.2201
y₄-NH₃514.262
b₅-H₂O528.2664
y₄531.2885
b₅546.277
y₅-NH₃627.3461
y₅644.3726
b₆-H₂O656.325
b₆674.3355
y₆-NH₃758.3865
b₇-H₂O769.409
y₆775.4131
b₇787.4196
y₇-NH₃871.4706
y₇888.4971
b₈-H₂O900.4495
y₈-H₂O998.5452
y₈-NH₃999.5292
y₈1016.556
y₉-H₂O1127.588
y₉-NH₃1128.572
y₉1145.598 y₁₀
1246.646
y₁₁-H₂O1329.683
y₁₁-NH₃1330.667
y₁₁1347.694
020
4060
80100
120
00.5
11.5
22.5
33.5
44.5
[1e5]
200 400 600 800 1000 1200
Raw File Scan Method Score m/zMT1131-minusCu-Set2-02 29313 FTMS; HCD 228.54 781.41
- T Lb₂
T
y₁₁
b₃T
y₁₀
E
y₉
b₅Q
y₈
b₆L
y₇
b₇M
y₆
L
y₅
V
y₄
E
y₃
Q
y₂
R
y₁
-
y₁-NH₃158.0924
y₁175.119
a₂191.1179
b₂219.1128
y₂-NH₃287.135
y₂304.1615
y₃403.23
y₄-NH₃483.2562
y₄500.2827
y₅-NH₃540.2776
y₅557.3042
y₆-NH₃641.3253
y₆658.3519
y₇-H₂O769.3839
y₇787.3945
020
4060
80100
120
00.2
0.40.6
0.81
1.2
[1e5]
100 200 300 400 500 600 700 800 900
Raw File Scan Method Score m/zMT1131-plusCu-Set2-02 13402 FTMS; HCD 77.07 503.25
- A Fb₂
E
y₇
T
y₆
G
y₅
P
y₄
V
y₃
E
y₂
R
y₁
-
a₂115.0866
y₁-NH₃130.0863
b₂143.0815
y₁147.1128
b₃-H₂O226.1186
b₃244.1292
y₂-H₂O258.1448
y₂276.1554
b₄-H₂O341.1456
y₃347.1925
b₄359.1561
y₄418.2296
y₅517.298
y₆588.3352
b₈-H₂O639.3097
y₇645.3566
y₈716.3937
y₉831.4207
y₁₀932.4684
020
4060
80100
120
00.5
11.5
22.5
3
[1e4]
200 400 600 800 1000
Raw File Scan Method Score m/zMT1131-plusCu-Set2-02 7165 FTMS; HCD 78.49 537.78
- A Ab₂
T
y₁₀
b₃D
y₉
b₄A
y₈
G
y₇
A
y₆
V
y₅
A
y₄
A
y₃
E
y₂
K
y₁
-
y₁175.119
a₂199.1805
b₂227.1754
b₃342.2023
b₅584.329
y₁₂²⁺653.8468
y₆673.3628
y₁₆²⁺845.9472
y₁₉²⁺1015.045
y₂₀²⁺1079.566
y₁₃1419.77
y₁₈1915.998
y₂₀2158.125
y₂₁2271.209
y₂₂2386.236
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
[1e5]
500 1000 1500 2000
Raw File Scan Method Score m/zMT1131-minusCu-Set1-02 33696 FTMS; HCD 80.16 871.81
- I Ib₂
D
y₂₂
b₃I
y₂₁
E
y₂₀
b₅L
y₁₉²⁺
P
y₁₈
Q G
y₁₆²⁺
V D L
y₁₃
P
y₁₂²⁺
R P A P D
A
y₆
Q A L D R
y₁
-
a₂157.1335
y₁-NH₃158.0924
y₁175.119
b₂185.1285
b₃313.187
b₄442.2296
y₅459.2674
b₅541.298
y₆558.3358
y₇-NH₃688.3777
y₇705.4042
b₇785.4192
y₈802.457 y₉
949.5254
y₁₀1048.594 y₁₁
1177.636
y₁₂-NH₃1288.668
y₁₂1305.695
020
4060
80100
120
01
23
45
6
[1e5]
200 400 600 800 1000 1200 1400 1600 1800
Raw File Scan Method Score m/zMT1131-plusCu-03 44457 FTMS; HCD 60 745.42
- A Lb₂
Q
y₁₂
b₃E
y₁₁
b₄V
y₁₀
b₅F
y₉
P
y₈
b₇F
y₇
V
y₆
A
y₅
A A A R
y₁
-
y₁-NH₃158.0924
y₁175.119
a₂187.1077
b₂215.1026
b₃312.1554
y₃-NH₃371.2037
y₃388.2303
y₄501.3144
y₅-NH₃571.3198
y₅588.3464
y₆-NH₃628.3413
y₆645.3678
y₇-H₂O698.3944
y₇716.405
y₈787.4421
y₉-H₂O866.4843
y₉-NH₃867.4683
y₉884.4948
020
4060
80100
120
00.2
0.40.6
0.81
1.2
[1e6]
200 400 600 800 1000 1200
Raw File Scan Method Score m/zMT1131-plusCu-01_150512102711 14088 FTMS; HCD 66.96 549.8
- V Db₂
P
y₉
b₃A
y₈
A
y₇
G
y₆
S
y₅
L
y₄
N
y₃
V R
y₁
-
y₁-NH₃158.0924
y₁175.119
b₂-NH₃227.1139
b₂244.1404
y₄625.3093
b₆669.3427
b₇-NH₃767.3431
b₁₄²⁺784.3478
b₇784.3696
y₆854.3791 y₈
1054.459y₁₀-NH₃
1205.522
y₁₀1222.549
b₁₃1381.609
y₁₅1762.778
020
4060
80100
120
00.5
11.5
22.5
33.5
44.5
5
[1e5]
200 400 600 800 1000 1200 1400 1600 1800 2000
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-01 13431 FTMS; HCD 55.55 669.64
- S Rb₂
P
y₁₅
G R Db₆
Db₇
P
y₁₀
A A
y₈
E N
y₆
Db₁₃
W
y₄
b₁₄²⁺Y T R
y₁
-
a₂131.0815
b₂-H₂O141.0659
y₁-NH₃158.0924
b₂159.0764
y₁175.119
b₃-H₂O212.103
b₃230.1135
b₄-H₂O311.1714
y₂-NH₃321.1557
b₄329.1819
y₂338.1823
y₃-NH₃408.1878
b₅-H₂O412.2191
y₃425.2143 y₄-NH₃
479.2249
y₄496.2514
y₅-H₂O575.2936
y₅-NH₃576.2776
y₅593.3042
y₆-H₂O662.3257
y₆-NH₃663.3097
y₆680.3362
y₇-H₂O763.3733
y₇781.3839
y₈-NH₃863.4258
y₈880.4523
y₉951.4894
y₁₀1008.511
020
4060
80100
120
00.5
11.5
22.5
33.5
[1e5]
200 400 600 800 1000 1200
Raw File Scan Method Score m/zMT1131-minusCu-03 10695 FTMS; HCD 137.28 555.28
- T G
y₁₀
b₂A
y₉
b₃V
y₈
b₄T
y₇
S
y₆
P
y₅
A
y₄
S
y₃
Y
y₂
R
y₁
-
a₂115.0866
y₁-NH₃130.0863
b₂143.0815
y₁147.1128
b₃214.1186
y₂218.1499
b₄285.1557
y₃305.1819
b₅382.2085
b₇578.3297
y₆-H₂O627.3097
y₆645.3202
y₇-H₂O756.3523
y₇774.3628
y₈845.3999
y₉942.4527
y₁₀1041.521
y₁₁1138.574
y₁₂1235.627
020
4060
80100
120
00.5
11.5
22.5
[1e5]
200 400 600 800 1000 1200
Raw File Scan Method Score m/zMT1131-minusCu-Set2-01 15756 FTMS; HCD 65.35 760.39
- A Ab₂
Ab₃
Ab₄
P
y₁₂
b₅P
y₁₁
V
y₁₀
b₇P
y₉
A
y₈
E
y₇
P
y₆
Q D S
y₃
A
y₂
K
y₁
-
y₁-NH₃158.0924
y₁175.119
y₂-NH₃215.1139
y₂232.1404
a₂233.1648
b₂261.1598
y₃345.2245 b₃
375.2027y₄
474.2671 y₆716.3937
y₇-H₂O769.4203
y₇787.4308
y₈-NH₃867.4571
y₈884.4836
y₁₆²⁺904.4705
y₁₇²⁺978.0047 y₉-H₂O
1003.532
y₉-NH₃1004.516
y₉1021.543
y₁₀1134.627
y₁₁1247.711
y₁₂1344.763
y₁₃-H₂O1455.795
y₁₃1473.806
y₁₄1530.827
y₁₅1693.891
y₁₆-NH₃1790.907
020
4060
80100
120
00.2
0.40.6
0.81
1.2
[1e6]
200 400 600 800 1000 1200 1400 1600 1800
Raw File Scan Method Score m/zMT1131-plusCu-01_150512102711 44789 FTMS; HCD 66.89 690.03
- L F
y₁₇²⁺
b₂N
y₁₆²⁺
b₃Y
y₁₅
G
y₁₄
E
y₁₃
P
y₁₂
L
y₁₁
L
y₁₀
H
y₉
P
y₈
A
y₇
L
y₆
E E
y₄
L
y₃
G
y₂
R
y₁
-
y₁-NH₃158.0924
a₂159.1128
b₂-H₂O169.0972
y₁175.119
b₂187.1077
y₂274.1874
b₃302.1347
b₄-H₂O385.1718
y₃-NH₃385.2194
y₃402.2459
b₄403.1823
y₄-NH₃484.2878
b₅-H₂O499.2147
b₅-NH₃500.1987
y₄501.3144
y₅602.362
b₆-H₂O646.2831
b₆664.2937
y₆-H₂O698.3944
y₆-NH₃699.3784
y₆716.405
y₇-H₂O845.4628
y₇-NH₃846.4468
y₇863.4734
y₈-NH₃960.4898
y₈977.5163
y₉-NH₃1061.537
y₉1078.564
y₁₀-H₂O1175.58
y₁₀-NH₃1176.564
y₁₀1193.591
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
[1e6]
200 400 600 800 1000 1200
Raw File Scan Method Score m/zMT1131-plusCu-01_150512102711 19544 FTMS; HCD 192.26 690.35
- S Vb₂
D
y₁₀
b₃T
y₉
b₄N
y₈
F
y₇
b₆N
y₆
T
y₅
V
y₄
Q
y₃
V
y₂
R
y₁
-
y₁-NH₃130.0863
y₁147.1128
a₂201.1234
b₂-H₂O211.1077
b₂229.1183
y₂248.1605
b₃-H₂O324.1918
b₃342.2023
y₃361.2445
b₄-H₂O395.2289
b₄413.2395 b₅-H₂O
466.266
y₄476.2715
y₅662.3508
y₆719.3723
y₇-H₂O798.4145
y₇816.425
y₈887.4621
y₉-H₂O940.4887
y₉958.4993
y₁₀1029.536
y₁₁1142.62
020
4060
80100
120
00.2
0.40.6
0.81
1.2
[1e5]
200 400 600 800 1000 1200 1400
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-03 31175 FTMS; HCD 71.9 685.87
- E Vb₂
L
y₁₁
b₃A
y₁₀
b₄A
y₉
A
y₈
P
y₇
G
y₆
W
y₅
D
y₄
L
y₃
T
y₂
K
y₁
-
b₂277.15
y₃-NH₃358.21
y₃375.24
y₄-H₂O454.28
y₄-NH₃455.26
y₄472.29
y₅571.36
b₅592.31
b₆-H₂O689.33
y₆700.4
b₇-H₂O788.4
b₇806.41
y₇829.44
b₈877.45
y₈916.47
b₉-H₂O946.47
b₉964.48
y₉987.51
b₁₀-H₂O1075.51
y₁₀1086.58
b₁₀1093.52
y₁₁1201.61
b₁₁1222.56
y₁₂1272.64
b₁₂-H₂O1303.62
b₁₂1321.63
y₁₃1419.71
y₁₄1516.76
b₁₄1531.77
b₁₅1618.80
2040
6080
100
120
00.5
11.5
22.5
3
[1e4]
400 600 800 1000 1200 1400 1600 1800
Raw File Scan Method Score m/zPlusCu-Biorep2-Run2 13179 ITMS; CID 164.48 897.46
- Y Lb₂
P
y₁₄
F
y₁₃
A
y₁₂
b₅D
y₁₁
V
y₁₀
b₇A
y₉
b₈S
y₈
b₉E
y₇
b₁₀E
y₆
b₁₁V
y₅
b₁₂P
y₄
L
y₃
b₁₄Sb₁₅
R -
a₂127.0866
b₂155.0815
y₁-NH₃158.0924
y₁175.119
b₃-H₂O224.103
y₂-NH₃229.1295
b₃242.1135
y₂246.1561
b₄-H₂O281.1244
b₄299.135
y₃-NH₃342.2136
y₃359.2401
b₅-H₂O382.1721
b₅400.1827
y₄-NH₃489.282
y₄506.3085
y₅619.3926
y₆-NH₃699.4188
y₆716.4454
y₇817.4931
y₈-H₂O856.5039
y₈874.5145
y₉-H₂O943.536
y₉-NH₃944.52
y₉961.5465
y₁₀-H₂O1000.557
y₁₀1018.568
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
1.61.8
[1e5]
200 400 600 800 1000
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-Set2-01 25672 FTMS; HCD 152.79 558.31
- P G
y₁₀
b₂S
y₉
b₃G
y₈
b₄T
y₇
b₅P
y₆
L
y₅
F
y₄
L
y₃
A
y₂
R
y₁
-
y₃-NH₃373.12
y₃390.14
y₄518.2
y₅617.27
y₆-NH₃701.29
y₆718.32
b₈-H₂O800.32
b₈818.33
y₈904.42
y₉991.45
b₁₁-H₂O1101.49
y₁₀1104.54
b₁₁1119.5
y₁₁-NH₃1188.56
y₁₁1205.58
b₁₂1206.53
y₁₂1292.62
b₁₃1293.56
b₁₄-H₂O1374.62
b₁₄1392.63
b₁₅1493.68
b₁₆-H₂O1574.74
b₁₆1592.75
y₁₅1607.76
b₁₇-H₂O1702.8
b₁₇-NH₃1703.78
b₁₇1720.81
b₁₈-H₂O1816.84
b₁₈-NH₃1817.82
b₁₈1834.85
020
4060
80100
120
0500
10001500
20002500
30003500
40004500
400 600 800 1000 1200 1400 1600 1800
Raw File Scan Method Score m/zPlusCu-Biorep2-Run3 11287 ITMS; CID 170 1055.98
- C E G G V S
y₁₅
D Lb₈
S
y₁₂
T
y₁₁
L
y₁₀
b₁₁S
y₉
b₁₂S
y₈
b₁₃Vb₁₄
T
y₆
b₁₅V
y₅
b₁₆Q
y₄
b₁₇N
y₃
b₁₈
P C -
b₂115.0502
y₁-NH₃158.0924
y₁175.119
b₃186.0873
y₂-NH₃321.1557
y₂338.1823
y₃-NH₃434.2398
y₃451.2663
y₄-NH₃571.2987
y₄588.3253
y₅-NH₃668.3515
y₅685.378
y₆-NH₃815.4199
y₆832.4464
020
4060
80100
120
01
23
45
67
[1e5]
200 400 600 800
Raw File Scan Method Score m/zMT1131-minusCu-01 23501 FTMS; HCD 76.83 377.54
- G Gb₂
Ab₃
L F
y₆
P
y₅
H
y₄
L
y₃
Y
y₂
R
y₁
-
a₂127.0866
b₂155.0815
y₁-NH₃158.0924
y₁175.119
b₃252.1343
y₂-NH₃273.1193
y₄448.215
y₅519.2522
y₆-H₂O558.2631
y₆576.2736
y₇673.3264
y₈786.4104
y₉-H₂O865.4526
y₉883.4632
y₁₀-NH₃1022.538
y₁₀1039.564
y₁₁1168.607
y₁₂-NH₃1222.617
y₁₂1239.644
y₁₃1310.681
y₁₄1407.734
y₁₅-NH₃1447.729
y₁₅1464.755
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
[1e5]
200 400 600 800 1000 1200 1400
Raw File Scan Method Score m/zMT1131-plusCu-Set2-01 12688 FTMS; HCD 68.58 521.27
- P G
y₁₅
b₂P
y₁₄
b₃A
y₁₃
A
y₁₂
E
y₁₁
R
y₁₀
P
y₉
L
y₈
P
y₇
G
y₆
A
y₅
S
y₄
A D R
y₁
-
y₁-NH₃158.0924
y₁175.119
b₂-H₂O213.087
b₂231.0975
y₂232.1404
b₃-H₂O314.1347
b₃332.1452
y₄460.2514
b₅-H₂O484.2402
y₅588.31
y₆717.3526
y₇830.4367
y₈977.5051 y₉
1124.573
y₁₀1195.611
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
1.6
[1e4]
200 400 600 800 1000 1200 1400 1600
Raw File Scan Method Score m/zMT1131-plusCu-Set3-02 41150 FTMS; HCD 72.65 813.41
- T Eb₂
Tb₃
V A
y₁₀
F
y₉
F
y₈
L
y₇
E
y₆
Q
y₅
L
y₄
D G
y₂
R
y₁
-
y₁-NH₃158.0924
b₂-NH₃169.0608
y₁175.119
b₂186.0873
b₃-NH₃240.0979
b₃257.1244
y₂-NH₃286.151
y₂303.1775
b₄-H₂O340.1615
y₃-NH₃357.1881
y₃374.2146
y₄503.2572
y₅560.2787
y₆631.3158
y₇762.3563
y₈863.404
y₉-H₂O916.4305
y₉934.4411
020
4060
80100
120
00.5
11.5
22.5
33.5
4
[1e4]
200 400 600 800 1000
Raw File Scan Method Score m/zMT1131-plusCu-Set3-01 5779 FTMS; HCD 101.65 560.26
- G Qb₂
A
y₉
b₃T
y₈
M
y₇
A
y₆
G
y₅
E
y₄
A
y₃
Q
y₂
R
y₁
-
y₁-NH₃158.0924
a₂173.0921
y₁175.119
b₂-H₂O183.0764
b₂201.087
b₃272.1241
y₂289.1619
b₄-H₂O369.1405
b₄387.151
y₃388.2303
y₇²⁺435.7489
b₅458.1882
y₅-NH₃569.3042
y₅586.3307
b₁₂²⁺620.3018
y₆-NH₃725.4053
y₁₃²⁺728.8606
y₇-NH₃853.4639
y₈-NH₃924.501
y₈941.5275
y₉-NH₃1053.544
y₉1070.57
y₁₀1141.607
y₁₁1256.634
y₁₂-H₂O1309.661
y₁₂1327.671
020
4060
80100
120
01
23
45
67
89
[1e4]
200 400 600 800 1000 1200
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-Set2-03 7237 FTMS; HCD 106.17 509.92
- A E
y₁₃²⁺
b₂A
y₁₂
b₃D
y₁₁
b₄A
y₁₀
b₅E
y₉
A
y₈
Q
y₇²⁺
R P
y₅
T V
y₃
b₁₂²⁺N
y₂
R
y₁
-
y₁-NH₃158.0924
y₁175.119
a₂201.1234
b₂229.1183
y₂-NH₃271.1765
y₂288.203
y₃-NH₃384.2605
y₃401.2871
y₄-NH₃512.3191
y₄529.3457
y₅642.4297
y₆729.4618
y₇-H₂O768.4726
y₇-NH₃769.4567
y₇786.4832
y₈-H₂O865.5254
y₈883.536
y₉998.56290
2040
6080
100
120
00.2
0.40.6
0.81
1.21.4
1.61.8
22.2
[1e5]
200 400 600 800 1000 1200
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-03 30374 FTMS; HCD 100.02 556.33
- L D
y₉
b₂P
y₈
G
y₇
S
y₆
L
y₅
Q
y₄
I
y₃
L
y₂
R
y₁
-
y₁-NH₃158.0924
y₁175.119
a₂193.0972 b₂
221.0921
y₂-NH₃229.1295
y₂246.1561
b₃335.135
y₃359.2401
b₄392.1565
y₄-NH₃470.2722
y₄487.2987
y₅-H₂O598.3307
y₅-NH₃599.3148
y₅616.3413
y₆-H₂O695.3835
y₆-NH₃696.3675
y₆713.3941
y₇-H₂O752.405
y₇-NH₃753.389
y₇770.4155
y₈-NH₃867.4319
y₈884.4585
y₉-NH₃924.4534
y₉941.4799
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
1.61.8
22.2
[1e5]
200 400 600 800 1000
Raw File Scan Method Score m/zMT1131-minusCu-Set2-02 11575 FTMS; HCD 141.23 552.78
- Y G
y₉
b₂N
y₈
b₃G
y₇
b₄P
y₆
E
y₅
Q
y₄
I
y₃
A
y₂
R
y₁
-
y₁116.0706
a₂145.0972
b₂-H₂O155.0815
b₂173.0921
y₂272.1717
y₃-H₂O383.2037
y₃-NH₃384.1878
y₆²⁺384.2298
y₃401.2143
y₄-H₂O480.2565
y₄-NH₃481.2405
y₄498.2671
y₅-NH₃637.3416
y₁₁²⁺664.8879
y₆-NH₃750.4257
y₇-NH₃863.5098
y₇880.5363
y₈-NH₃976.5938
y₈993.6204 y₉
1156.684
y₁₀1257.731
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
1.61.8
2
[1e5]
200 400 600 800 1000 1200
Raw File Scan Method Score m/zMT1131-minusCu-01 23330 FTMS; HCD 54.77 477.29
- T A
y₁₁²⁺
b₂T
y₁₀
Y
y₉
I
y₈
L
y₇
L
y₆²⁺
R P
y₄
E
y₃
R
y₂
P
y₁
-
y₁-NH₃130.0863
y₁147.1128
y₂-H₂O216.1343
a₂217.1335
y₂234.1448
b₂245.1285
b₃316.1656
y₃381.2132
b₄387.2027
y₄494.2973
b₆-H₂O607.2987
y₅607.3814
b₆625.3093
y₆694.4134
b₇-H₂O720.3828
b₇738.3933
y₇807.4975
b₈835.4461 y₈
864.5189
b₉-H₂O904.4676
b₉922.4781
y₉-H₂O943.5611
y₉961.5717
b₁₀-H₂O1051.536
b₁₀1069.547
y₁₀1074.656
b₁₁-H₂O1108.557
b₁₁1126.568
b₁₂-H₂O1237.6
b₁₂1255.611
b₁₃1368.695
y₁₅1591.8730
2040
6080
100
120
00.2
0.40.6
0.81
1.21.4
1.6
[1e6]
200 400 600 800 1000 1200 1400 1600
Raw File Scan Method Score m/zMT1131-plusCu-01_150512102711 61107 FTMS; HCD 91.28 777.43
- F Pb₂
Ab₃
Ab₄
T Hb₆
Lb₇
P
y₁₅
b₈Sb₉
Fb₁₀
Gb₁₁
Eb₁₂
I
y₁₀
b₁₃P
y₉
G
y₈
L
y₇
S
y₆
L
y₅
I
y₄
F
y₃
S
y₂
K
y₁
-
y₁-NH₃158.0924
y₁175.119
a₂199.1805
b₂227.1754
y₂288.203
b₃298.2125
y₃359.2401
y₄-NH₃457.2405
y₄474.2671
y₉²⁺500.2375
y₅-NH₃528.2776
y₅545.3042
y₁₂²⁺671.8064
y₆676.3447
y₁₄²⁺771.8645
y₇-NH₃796.377
y₇813.4036
y₈928.4305
y₉999.4676
y₁₀-NH₃1097.468
y₁₀1114.495
y₁₁-H₂O1211.511
y₁₁1229.522
y₁₂-H₂O1324.595
y₁₂1342.606
y₁₃1455.69
y₁₄-H₂O1524.711
y₁₄1542.722
y₁₅-H₂O1671.78
y₁₅1689.79
y₁₆1790.838
y₁₇-NH₃1886.895
y₁₇1903.922
y₁₈-H₂O1982.964
y₁₈2000.975
y₁₉2072.012
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
1.61.8
22.2
[1e5]
200 400 600 800 1000 1200 1400 1600 1800 2000
Raw File Scan Method Score m/zMT1131-plusCu-02 47093 FTMS; HCD 150.69 767.07
- L Lb₂
A
y₁₉
b₃P
y₁₈
L
y₁₇
T
y₁₆
F
y₁₅
S
y₁₄
L
y₁₃
L
y₁₂
D
y₁₁
D
y₁₀
A
y₉
D
y₈
H
y₇
M
y₆
A
y₅
D
y₄
A
y₃
L
y₂
R
y₁
-
y₁-NH₃158.0924
b₂-NH₃169.0608
y₁175.119
b₂186.0873
b₃-NH₃266.1135
b₃283.1401
y₂-NH₃321.1557
y₂338.1823
b₄-H₂O380.1565
b₄-NH₃381.1405
y₃-NH₃449.2143
y₃466.2409
b₅-H₂O479.2249
b₆-H₂O536.2463
y₄-NH₃546.2671
y₄563.2936
y₅-NH₃603.2885
y₅620.3151
y₆-NH₃702.357
y₆719.3835
y₇-NH₃817.3839
y₇834.4104
y₈931.4632
y₉988.4847
020
4060
80100
120
00.2
0.40.6
0.81
1.21.4
1.61.8
22.2
[1e5]
200 400 600 800 1000
Raw File Scan Method Score m/zMT1131-minusCu-Set3-01 12049 FTMS; HCD 88.92 558.78
- Q G
y₉
b₂P
y₈
b₃D
y₇
V
y₆
G
y₅
P
y₄
Q
y₃
Y
y₂
R
y₁
-
a₂158.0924
b₂-NH₃169.0608
b₂186.0873
y₂-H₂O225.0982
y₂243.1088
b₃-NH₃282.1448
b₃299.1714
y₃-H₂O388.1615
b₄-H₂O396.1878
b₄-NH₃397.1718
y₃406.1721 b₄
414.1983
y₄-H₂O459.1987
b₅-NH₃510.2558
y₅564.2413
b₆-NH₃611.3035
y₇-H₂O732.2947
y₇750.3053
b₈-NH₃849.4101
y₈887.3642
b₉-NH₃906.4316
y₉-H₂O970.4013
y₉988.4119
b₁₀-H₂O1034.49
b₁₀-NH₃1035.474
y₁₀-H₂O1071.449
y₁₀1089.46
b₁₁-H₂O1121.522
b₁₂-NH₃1193.543
y₁₁1202.544
b₁₂1210.57
y₁₂1317.571
b₁₃-H₂O1355.623
b₁₃1373.633
020
4060
80100
120
00.2
0.40.6
0.81
[1e5]
200 400 600 800 1000 1200 1400 1600
Raw File Scan Method Score m/zMT1131-minusCu-Set2-02 18113 FTMS; HCD 160.53 808.37
- Q Gb₂
Lb₃
D
y₁₂
b₄L
y₁₁
T
y₁₀
T
y₉
H
y₈
G
y₇
E S
y₅
Ab₁₂
Y
y₃
b₁₃H
y₂
S -
y₁133.0608
y₂-H₂O244.0928
a₂272.1757
b₂300.1707
y₃-NH₃359.1197
y₃376.1463 b₃
415.1976
b₄571.2987
b₅-H₂O668.3151
b₅-NH₃669.2991
b₅686.3257
y₆-NH₃743.3319
y₆760.3584
b₆799.4097
y₇-H₂O857.3748
y₇-NH₃858.3588
y₇875.3854
b₇-NH₃896.4261
b₇913.4526
b₈1042.495
y₈-H₂O1043.454
y₈-NH₃1044.438
y₈1061.465
020
4060
80100
120
00.2
0.40.6
0.81
[1e5]
200 400 600 800 1000
Raw File Scan Method Score m/zMT1131-minusCu-Set2-02 16809 FTMS; HCD 104.01 587.78
- I W
y₈
b₂D
y₇
b₃R
y₆
b₄Db₅
Ib₆
N
y₃
b₇Eb₈
N
y₁
-
b₂145.0608
y₁-NH₃158.0924
y₁175.119
b₃274.1034
y₂288.203
b₄403.146
y₃403.23 y₄
474.2671
b₅531.2045
y₅-H₂O585.2991
y₅603.3097
b₆630.2729
y₆716.3937
y₇-NH₃796.4199
y₇813.4465
b₈824.3533
y₈-H₂O923.4945
y₈-NH₃924.4785
y₈941.5051
b₉971.4217
y₉-NH₃981.5
y₉998.5265
y₁₀-NH₃1052.537
b₁₀-H₂O1054.459
y₁₀1069.564
b₁₀1072.469
b₁₁-H₂O1167.543
y₁₁1182.648
b₁₁1185.553
b₁₂1298.638
020
4060
80100
120
00.5
11.5
22.5
33.5
[1e5]
200 400 600 800 1000 1200 1400
Raw File Scan Method Score m/zMT1131-plusCu-Set2-01 33816 FTMS; HCD 80.31 790.07
- S Gb₂
Eb₃
Eb₄
Qb₅
Vb₆
G Hb₈
Fb₉
Tb₁₀
Lb₁₁
I
y₁₁
b₁₂A
y₁₀
G
y₉
Q
y₈
P
y₇
L
y₆
E
y₅
A
y₄
D
y₃
L
y₂
R
y₁
-
a₂101.0709
b₂129.0659
y₁-NH₃158.0924
y₁175.119
b₃242.1499
y₂-NH₃286.151
y₂303.1775
y₃-NH₃343.1724
y₃360.199
y₄-NH₃414.2096
y₄431.2361
y₅-NH₃501.2416
y₅518.2681
y₆-NH₃600.31
y₆617.3365
y₇-NH₃687.342
y₇704.3686
y₈-H₂O787.4057
y₈805.4163
020
4060
80100
120
01
23
45
67
[1e4]
200 400 600 800 1000
Raw File Scan Method Score m/zMT1131-minusCu-plusBC-02 9019 FTMS; HCD 104.06 523.78
- A Gb₂
Lb₃
T
y₈
S
y₇
V
y₆
S
y₅
A
y₄
G
y₃
Q
y₂
R
y₁
-
y₁175.119
a₂205.1005
b₂233.0954
y₂288.203
b₃346.1795
y₃389.2507
b₄-H₂O456.2275
b₄474.2381
y₄504.2776
b₁₀²⁺554.7582
b₁₁²⁺628.2924
b₁₂²⁺701.8266
y₆702.3781
y₇773.4152
y₈844.4523
b₈922.4451
y₉941.5051
y₁₀1012.542
b₉1037.472
b₁₀1108.509
y₁₁1127.569
y₁₂1240.653
b₁₁1255.578
b₁₂1402.646
b₁₃-NH₃1456.657
b₁₃1473.683
b₁₆1772.831
b₁₇1843.868
020
4060
80100
120
01
23
45
6
[1e5]
200 400 600 800 1000 1200 1400 1600 1800
Raw File Scan Method Score m/zMT1131-minusCu-03 44031 FTMS; HCD 86.13 928.79
- T Mb₂
Lb₃
Qb₄
P V D Hb₈
Db₉
Ab₁₀
Fb₁₁
Fb₁₂
Ab₁₃
A L
y₁₂
D
y₁₁
b₁₆A
y₁₀
b₁₇P
y₉
A
y₈
A
y₇
P
y₆
T D
y₄
T
y₃
L
y₂
R
y₁
-
S- 15
Figure S5. Total cellular metal contents of cells grown in Cu-sufficient control (-Cu), or + Cu (5
µM), or +BCS (5 mM) medium, determined by ICP-MS as described earlier (8, 9) (note the
logarithmic scale of y axis).
Figure S5. Selamoglu et al.