Table 10.
Identification and quantification of enzymes and other factors by 2DE-MS/MS.
| Spot # | Predominant protein1 | Total proteins in spot2 |
Spot Volume3 |
Std Dev4 |
MS/MS Coverage5 |
Predicted MW6 |
pI7 |
|---|---|---|---|---|---|---|---|
| Beta-amylase (β-Amy-A1, β-Amy-B1, β-Amy-D1) | |||||||
| 476 | Beta-amylase Bu-18 | 4 | - | - | 32 | 60016 | 6.9 |
| 537 | Beta-amylase Bu-1 | 2 | 0.04 | 0.010 | 18 | 60016 | 6.9 |
| 94 | Beta-amylase Bu2 | 2 | 0.04 | 0.006 | 47 | 54481 | 5.9 |
| 462 | Beta-amylase Bu2 | 2 | 0.11 | 0.014 | 60 | 54481 | 5.9 |
| 64 | Beta-amylase Bu3 | 1 | 0.05 | 0.010 | 13 | 54319 | 5.8 |
| 92 | Beta-amylase Bu3 | 1 | 0.18 | 0.029 | 34 | 54319 | 5.8 |
| 93 | Beta-amylase Bu3 | 1 | 0.09 | 0.013 | 55 | 54319 | 5.8 |
| Other enzymes9 | |||||||
| 89 | ADP-glucose PP lg subunit [GenBank:CAD98749] | 1 | 0.03 | 0.002 | 10 | 53030 | 5.6 |
| 128 | ADP-glucose PP lg subunit [GenBank:CAD98749] | 1 | 0.10 | 0.047 | 11 | 53030 | 5.6 |
| 108 | ADP-glucose PP sm subunit [GenBank:AAF61173] | 2 | 0.09 | 0.013 | 60 | 52061 | 5.5 |
| 110 | ADP-glucose PP sm subunit [GenBank:AAF61173] | 2 | 0.06 | 0.019 | 24 | 52061 | 5.5 |
| 118 | Alanine amino transferase TC11_282456 | 1 | 0.06 | 0.002 | 36 | 52820 | 6.1 |
| 109 | ATP-synthase beta-subunit [GenBank:CAA52636] | 1 | 0.02 | 0.004 | 32 | 58562 | 5.4 |
| 232 | Chitinase, rye, [GenBank:BAB18520] | 1 | 0.04 | 0.009 | 40 | 26095 | 8.7 |
| 455 | Chitinase [GenBank:AAX83262] | 1 | 0.05 | 0.010 | 26 | 26022 | 8.3 |
| 241 | Dehydroascorbate reductase TC264934 | 2 | 0.09 | 0.013 | 83 | 23358 | 5.9 |
| 123 | Enolase TC11_292359 | 2 | 0.07 | 0.014 | 64 | 48033 | 5.4 |
| 178 | Glyoxalase I TC11_288238 | 1 | 0.10 | 0.020 | 68 | 32568 | 5.4 |
| 202 | Glucose/ribitol dehydrogenase RS_UWI_14903 | 1 | <0.01 | 0.000 | 57 | 31851 | 6.3 |
| 436 | Ketol-acid reducto isomerase TC234371 | 3 | 0.05 | 0.005 | 21 | 57486 | 5.4 |
| 175 | Malate dehydrogenase [GenBank:AAT64932] | 1 | 0.03 | 0.007 | 41 | 35486 | 5.8 |
| 176 | Malate dehydrogenase [GenBank:AAT64932] | 1 | 0.15 | 0.000 | 58 | 35486 | 5.8 |
| 450 | Methionine synthase RS_UWI_10957 | 1 | 0.02 | 0.005 | 11 | 84552 | 5.7 |
| 371 | Orthophosphate dikinase TC11_322894 | 1 | 0.03 | 0.004 | 19 | 73501 | 5.8 |
| 299 | PDI3 [GenBank:AAK49425] | 3 | 0.14 | 0.016 | 53 | 54094 | 4.9 |
| 475 | Sucrose synthase 2 [GenBank:CAA03935] | 1 | 0.08 | 0.011 | 19 | 92608 | 6.2 |
| 189 | Thiamine biosynth enzyme TC11_308909 | 2 | 0.08 | 0.007 | 35 | 33167 | 5.7 |
| 225 | Triose-phosphate isomerase [GenBank:CAC14917] | 1 | 0.05 | 0.006 | 38 | 26803 | 5.4 |
| 239 | 27 K thiol reductase-like TC11_300123 | 1 | 0.16 | 0.037 | 54 | 23642 | 6.1 |
| 479 | 27 K thiol reductase like TC11_299048 | 2 | 0.02 | 0.005 | 16 | 23788 | 6.1 |
| Other | |||||||
| 311 | Elongation factor EF1A [Swiss-Prot: Q03033] | 1 | 0.11 | 0.005 | 14 | 49169 | 9.2 |
| 533 | HSP70 (Butte 86) [GenBank:AAB99745] | 1 | 0.08 | 0.009 | 18 | 71031 | 5.1 |
| 413 | Initiation factor Eif4A [Swiss-Prot: P41378.1] | 1 | 0.06 | 0.014 | 36 | 46928 | 5.3 |
| 295 | LTP Bu-2 | 1 | 0.04 | 0.009 | 38 | 9606 | 8.2 |
| 456 | Thaumatin-like protein TC11_283136 | 2 | 0.05 | 0.007 | 39 | 21408 | 7.9 |
Loci are indicated within parentheses if known.
1 Protein that matched the greatest number of peptides detected in spot by MS/MS. Accession numbers from TaGI Release 10.0 are denoted with the prefix TC and TaGI release 11.0 with the prefix TC11_. Accession numbers from HarvEST 1.14 are denoted with RS_UWI_. Sequences of proteins from Butte 86 are found in Additional file 2.
2-7 Footnotes as in Table 3.
8Although Scaffold identified more peptides for beta-amylase in spot 476, other data indicate that this spot was predominantly omega-gliadin.
9The following enzymes or factors were detected as minor components of other spots: alcohol dehydrogenase, aspartic protease, formate dehydrogenase, fructose bisphosphate aldolase, GDP-binding protein, manganese superoxide dismutase, peroxidase, peroxiredoxin, phosphoglycerate kinase.