Table 1.
Redox responsitive proteins identified in B. napus guard cells under ABA treatment.a
| Name | Accession (gi) | Mr (kDa)/pI | Unused Score (ICAT) | Peptide (ICAT) | Fold Change (ICAT) | Mascot Score (DIGE) | Spot No./Fold Change | iTRAQ ratio |
|---|---|---|---|---|---|---|---|---|
| Photosynthesis (6) | ||||||||
| Photosystem II 44 kDa reaction center | 131285 | 63/6.7 | 130 | 1093/0.36 | 1.20S | |||
| Ribulose-5-phosphate kinase | 21839 | 45/5.8 | 135 | 1546/1.85 | 0.92NS | |||
| &Ribulose bisphosphate carboxylase large chain | 1346967 | 62/6.2 | 27.81 | GRPLLGCTIKPK | 1.54 | 0.47S | ||
| CYHIEPVPGEETQFIAY | 1.46 | |||||||
| VALEACVQAR | 1.45 | |||||||
| WSPELAAACEVWK | 1.29 | |||||||
| YGRPLLGCTIKPK | 0.77 | |||||||
| GHYLNATAGTCEEMMK | 0.75 | |||||||
| &Sedoheptulose-bisphosphatase (SBPASE) | 15228194 | 43/6.2 | 208 | 1767/1.75 | 0.60S | |||
| &Rubisco small subunit | 17852 | 20/8.2 | 8.55 | WIPCVEFELEHGFVYR | 1.38 | 181 | 1093/0.36 | 0.97NS |
| Ferredoxin | 119980 | 10/3.8 | 2.03 | FITPEGEQEVECDDDVYVLDAAEEAGIDLPYSCR | 1.31 | NO ID | ||
| Energy (19) | ||||||||
| &De-etiolated 3, V-ATPase subunit C | 18391442 | 43/5.4 | 130 | 1546/1.85 | 1.08NS | |||
| &Putative fructose bisphosphate aldolase | 14334740 | 43/6.5 | 93 | 1762/UC | 1.32NS | |||
| Cytosolic triosephosphatisomerase | 15233272 | 27/5.4 | 7.72 | IIYGGSVNGGNCK | 1.46 | 118 | 2226/0.25 | 0.92S |
| &Fructose-bisphosphate aldolase | 14334740 | 43/4.4 | 342 | 1581/0.50 | 1.02NS | |||
| 43/5.8 | 238 | 1550/2.20 | ||||||
| 43/6.4 | 216 | 1577/0.46 | ||||||
| 43/5.0 | 175 | 1545/1.87 | ||||||
| 43/5.4 | 142 | 1546/1.85 | ||||||
| 43/5.3 | 128 | 1544/0.46 | ||||||
| &Chloroplast NAD-dependent malate dehydrogenase | 3256066 | 33/8.5 | 164 | 2051/1.67 | 1.01NS | |||
| Putative fructokinase | 14423528 | 35/5.3 | 123 | 1913/0.50 | 1.54NS | |||
| Transitional endoplasmic reticulum ATPase | 15232776 | 90/5.1 | 4.46 | QSAPCVLFFDELDSIATQR | 1.24 | 0.92NS | ||
| Mitochondrial malate dehydrogenase | 7769871 | 33/8.5 | 86 | 2016/3.06 | 1.16NS | |||
| ATP synthase subunit beta, mitochondrial | 114420 | 59/6.0 | 139 | 1223/1.87 | 1.24S | |||
| &F1-ATPase alpha subunit | 56384657 | 47/5.6 | 220 | 1107/0.29 | 0.69S | |||
| ATP synthase gamma chain, chloroplast | 5708095 | 33/6.1 | 112 | 1734/UC | 0.91NS | |||
| &Glyceraldehyde-3-phosphate dehydrogenase B precursor, chloroplast | 81621 | 43/5.6 | 220 | 1581/0.50 | 0.75S | |||
| &Malate dehydrogenase, mitochondrial | 899226 | 36/8.8 | 10.32 | GLNGVPDVVECSYVQSTITELPFFASK | 0.80 | 66 | 1975/0.41 | 1.37S |
| Succinate dehydrogenase flavoprotein | 15240075 | 69/5.9 | 4.49 | AAIGLSEHGFNTACITK | 0.76 | 192 | 794/0.53 | 1.40NS |
| &Malate dehydrogenase, cytosolic | 15219721 | 36/6.1 | 246 | 1795/0.25 | 0.66S | |||
| 36/4.8 | 111 | 1713/2.02 | ||||||
| &Phosphoglycerate kinase 1 (PGK1) | 15230595 | 43/5.3 | 173 | 1544/0.46 | 1.31S | |||
| &Vacuolar ATP synthase subunit A (VHA-A) | 15219234 | 69/4.8 | 6.01 | YSNSDAVVYVGCGER | 11.58 | 273 | 663/7.45 | 0.70S |
| Adenosine triphosphatase | 904041 | 54/5.0 | 130 | 1223/1.87 | NO ID | |||
| &Glyceraldehyde-3-phosphate dehydrogenase C subunit (GapC) | 15229231 | 37/6.6 | 8.04 | SDLDIVSNASCTTNCLAPLAK | 1.29 | NQ | ||
| Metabolism (12) | ||||||||
| Lactoylglutathione lyase | 2494843 | 32/5.2 | 58 | 2111/2.01 | 1.12NS | |||
| &3-ketoacyl-acyl carrier protein synthase I | 780814 | 51/8.0 | 52 | 1431/3.47 | 1.10NS | |||
| Enoyl-[acyl-carrier-protein] reductase | 99805 | 33/8.9 | 67 | 2016/3.06 | 1.08NS | |||
| Glycolate oxidase | 2501812 | 35/9.5 | 84 | 1762/UC | 0.78NS | |||
| &Reversibly glycosylated polypeptide-1 | 15232865 | 41/5.6 | 7.31 | NLLCPSTPFFFNTLYDPYR | 1.40 | 0.91NS | ||
| &Adenosine kinase 1 (ADK1) | 15232763 | 39/5.3 | 59 | 1762/UC | 1.18S | |||
| Glutamine synthetase | 6966930 | 62/6.4 | 83 | 1093/0.36 | 0.86NS | |||
| Dihydrodipicolinate reductase | 18406430 | 35/6.0 | 70 | 1913/0.50 | NO ID | |||
| Cinnamyl alcohol dehydrogenase | 1143445 | 43/8.2 | 56 | 1767/1.75 | NO ID | |||
| Biotin carboxyl carrier protein | 1070000 | 33/4.6 | 74 | 2106/UC | NO ID | |||
| Threonine synthase | 15233723 | 58/7.1 | 2.38 | HCGISHTGSFK | 0.66 | NO ID | ||
| &Oxalic acid oxidase | 60686421 | 22/9.1 | 9.45 | SVQDFCVANLKR | 1.90 | NO ID | ||
| AETPAGYPCIRPIHVK | 1.82 | |||||||
| Protein synthesis (5) | ||||||||
| 60S ribosomal protein L2 | 15227954 | 28/10.9 | 2.00 | SIPEGAVVCNVEHHVGDR | 0.57 | 1.08NS | ||
| Mitochondrial elongation factor Tu | 15236220 | 49/6.2 | 4.06 | QVGVPSLVCFLNK | 1.38 | 0.96NS | ||
| &Initiation factor 5A-4 | 15222741 | 17/5.6 | 2.01 | KLEDIVPSSHNCDVPHVNR | 1.23 | NO ID | ||
| Hypothetical protein, containing (EF1) domain | 147801436 | 58/6.7 | 74 | 1421/0.67 | NO ID | |||
| &Eukaryotic initiation factor 4A-2 | 1170506 | 47/5.9 | 95 | 1351/UC | NO ID | |||
| Protein folding, transporting and degradation (4) | ||||||||
| &Mitochondrial processing peptidase alpha subunit | 15218090 | 47/5.9 | 81 | 1351/UC | 0.98NS | |||
| &Putative aspartic protease | 510880 | 28/5.4 | 51 | 2061/1.67 | 0.97NS | |||
| Putative proteasome 20S beta1 subunit | 41352683 | 25/5.8 | 110 | 2434/3.09 | NO ID | |||
| Ubiquitin extension protein (UBQ5) | 15228715 | 18/9.8 | 1.52 | CGLTYVYQK | 0.64 | NO ID | ||
| Stress and defense (5) | ||||||||
| &Senescence-associated cysteine protease | 18141281 | 25/4.6 | 92 | 2426/2.55 | 1.19NS | |||
| &Low expression of osmotically responsive genes 1 (LOS2) | 15227987 | 48/5.5 | 127 | 1223/1.87 | 1.00NS | |||
| Early response to dehydration (ERD12) | 157849770 | 29/9.3 | 2.02 | NPQQLCIGDLVPFTNK | 1.29 | 0.74S | ||
| &Myrosinase, thioglucoside glucohydrolase | 414103 | 75/5.0 | 269 | 889/0.56 | 1.18S | |||
| 75/6.1 | 199 | 979/2.80 | ||||||
| 75/5.7 | 165 | 804/0.33 | ||||||
| 75/5.6 | 94 | 800/0.27 | ||||||
| 75/5.5 | 90 | 794/0.53 | ||||||
| Stromal ascorbate peroxidase | 46093471 | 38/7.1 | 2.80 | VDTSGPHECPEEGRLPDAGPPSPANHLR | 1.67 | 0.99NS | ||
| Signal transduction (4) | ||||||||
| Osmotic stress-activated protein kinase | 19568098 | 47/5.6 | 63 | 1351/UC | 0.99NS | |||
| 14-3-3 protein homolog | 100554 | 29/4.8 | 48 | 2111/2.01 | 0.86NS | |||
| Serine/threonine phosphatases 2C (PP2C) | 115468776 | 19/5.3 | 51 | 2630/2.89 | NO ID | |||
| Calmodulin-binding protein | 15242603 | 21/4.9 | 65 | 2546/2.66 | NO ID | |||
| Cell structure (5) | ||||||||
| &Actin | 4139264 | 43/5.4 | 298 | 1546/1.85 | 1.60NS | |||
| 43/4.5 | 220 | 1581/0.50 | ||||||
| 43/5.8 | 193 | 1550/2.20 | ||||||
| &Tubulin beta-4 chain(TUB4) | 15241472 | 63/4.8 | 217 | 1237/UC | NQ | |||
| &Plastid-lipid associated protein PAP2 | 14248550 | 25/4.8 | 54 | 2426/2.55 | NO ID | |||
| Putative protein, containing band 7 stomatin domain | 4469009 | 56/5.2 | 75 | 1577/0.46 | NO ID | |||
| &Extensin-like protein | 15235668 | 82/6.5 | 2.00 | IPASICQLPK | 1.53 | NO ID | ||
| Transcription (1) | ||||||||
| Retrotransposon protein, putative | 62733113 | 92/7.2 | 49 | 663/7.45 | NO ID | |||
| Cell division, differentiation and fate (3) | ||||||||
| Cell division protein FtsH | 15238333 | 75/5.4 | 3.12 | GCLLVGPPGTGK | 0.79 | 1.51S | ||
| GTP-binding nuclear protein RAN1 | 585777 | 29/6.2 | 76 | 2111/2.01 | NO ID | |||
| &Proliferating cell nuclear antigen (PCNA) | 2499441 | 33/4.6 | 62 | 2106/UC | NO ID | |||
| Unknown(1) | ||||||||
| Unnamed protein product | 134273558 | 52/6.8 | 2.19 | LLICGGSAYPR | 1.26 | 0.82S | ||
a
Overlapping proteins with MeJA results shown in Table 2 are labeled with ‘&’. Protein names in bold are newly identified as potentially redox regulated, and those in italics showed redox changes but were not identified or quantified in the iTRAQ experiments. Mr, molecular mass; pI, isoelectric point; ‘Spot No./Fold Change’, DIGE spot number and fold change of spot volume; UC, unique spot in control gel; ‘iTRAQ ratio’: NS, non-signficant quantification; S, significant quantification; NO ID, no identification; NQ, no quantification.