Table 2. Oxidation state of peptides showing a significant increase in cysteine S-nitrosylation with IPC as identified via SNO-RAC/Ox-RAC proteomic analysis.
| Protein Name | Protein ID | Peptide Sequence | SNO or Ox Cys | SNO Ratio (IPC/Perfusion) | Ox Ratio (IR/IPC-IR) |
|---|---|---|---|---|---|
| Aconitate hydratase (M) | Q99KI0 | VGLIGSCTNSSYEDMGR | 385 | 1.69† | |
| Aspartate aminotransferase | P05201 | INMCGLTTK | 391 | Not Detected in IR or IPC-IR | |
| Aspartate aminotransferase (M) | P05202 | VGAFTVVCK | 295 | 1.41† | |
| β-enolase | P21550 | VNQIGSVTESIQACK | 357 | 1.40† | |
| Cysteine and glycine-rich protein 3 | P50462 | TVYHAEEIQCNGR | 25 | 3.95† | |
| Cysteine-rich protein 2 | Q9DCT8 | ASSVTTFTGEPNMCPR | 126 | 1.70† | |
| Cytochrome b-c1 complex subunit 1 (M) | Q9CZ13 | VYEEDAVPGLTPCR | 268 | 1.57 | |
| LCTSATESEVTR | 380 | 7.31* | 0.67 | ||
| Cytochrome c oxidase subunit 5B (M) | P19536 | cPNCGTHYK | 115 | Not Detected in IPC-IR | |
| Electron transfer flavoprotein subunit α (M) | Q99LC5 | TIYAGNALCTVK | 155 | 1.55 | |
| Electron transfer flavoprotein subunit β (M) | Q9DCW4 | EIIAVSCGPSQCQETIR | 66,71 | 1.60† | |
| Enoyl-CoA hydratase (M) | Q8BH95 | LVEEAIQCAEK | 225 | 2.97† | |
| Fructose-bisphosphate aldolase A | P05064 | ALANSLACQGK | 339 | 9.90* | 2.04 |
| Glyceraldehyde-3-phosphate dehydrogenase | P16858 | IVSNASCTTNCLAPLAK | 150,154 | 2.97* | 2.74† |
| VPTPNVSVVDLTCR | 245 | 2.10* | 1.78† | ||
| Isocitrate dehydrogenase [NAD] subunit α (M) | Q9D6R2 | IEAACFATIK | 331 | Not Detected in IPC-IR | |
| Isocitrate dehydrogenase [NADP] (M) | P54071 | SSGGFVWACK | 308 | 1.60 | |
| VCVQTVESGAMTK | 402 | 1.91 | 0.99 | ||
| L-lactate dehydrogenase A chain | P06151 | IVSSKDYCVTANSK | 84 | 1.97 | |
| VIGSGCNLDSAR | 163 | 3.81* | 2.16 | ||
| Long-chain-fatty-acid-CoA ligase 1 | P41216 | GIQVSNNGPCLGSR | 109 | Not Detected in IR or IPC-IR | |
| Metallothionein-1 | P02802 | SCCSCCPVGCSK | 33,34,36 37,41 | Not Detected in IR or IPC-IR | |
| GAADKCTCCA | 57,59,60 | Not Detected in IR or IPC-IR | |||
| NADH dehydrogenase [ubiquinone] 1 α subcomplex subunit 10 (M) | Q99LC3 | VITVDGNICSGK | 67 | 1.69† | |
| Peroxiredoxin-6 | O08709 | DFTPVCTTELGR | 47 | 1.60† | |
| Propionyl-CoA carboxylase α chain | Q91ZA3 | MADEAVCVGPAPTSK | 107 | Not Detected in IR or IPC-IR | |
| Protein NipSnap homolog 2 | O55126 | ICQEVLPK | 89 | 22.40* | Not Detected in IR or IPC-IR |
| Succinate dehydrogenase [ubiquinone] flavoprotein (M) | Q8K2B3 | VGSVLQEGCEK | 536 | Not Detected in IR or IPC-IR | |
| Succinyl-CoA ligase subunit β (M) | Q9Z2I9 | ILACDDLDEAAK | 430 | 1.58† | |
| Triosephosphate isomerase | P17751 | IAVAAQNCYK | 67 | 3.40* | 5.65† |
| IIYGGSVTGATCK | 218 | 4.12† | |||
| Tubulin β-4 chain | P04350 | NMMAACDPR | 303 | 5.74† | |
| TAVCDIPPR | 354 | 1.54 | |||
| Voltage-dependent anion channel protein 2 (M) | Q60930 | SCSGVEFSTSGSSNTDTGK | 48 | 2.72† |
Each protein/peptide was identified from at least three of five SNO/Ox-RAC/LC-MS/MS proteomic analyses (peptides were filtered at a false discovery rate of 5%; peptides with ion scores below 30 were not accepted). (M): mitochondrial isoform; (C): SNO cysteine residue; (c): NEM-blocked cysteine residue; Peptides not detected under perfusion conditions contain a blank space in the ratio column.
*
p<0.05 Perfusion vs. IPC;
†
p<0.05 IR vs. IPC-IR.