Table 7.
Identification and quantification of alpha-gliadins by 2DE-MS/MS.
| Spot # | Predominant protein1 | Total proteins in spot2 |
Spot Vol3 |
Std Dev4 |
MS/MS Coverage5,8 |
Predicted MW6,8 |
pI7,8 | Celiac Peptides9 |
|---|---|---|---|---|---|---|---|---|
| Alpha-gliadins (Gli-A2) | ||||||||
| 183 | Alpha-gliadin Bu-5 | 3 | 1.23 | 0.076 | 76 | 30506 | 7.1 | 6 |
| 344 | Alpha-gliadin Bu-5 | 1 | 0.16 | 0.028 | 57 | 30506 | 7.1 | 6 |
| 331 | Alpha-gliadin Bu-14 | 2 | 0.28 | 0.041 | 15 | 29995 | 6.2 | 6 |
| 190 | Alpha-gliadin Bu-14 | 2 | 0.48 | 0.035 | 78 | 29995 | 6.2 | 6 |
| 206 | Alpha-gliadin Bu-14 | 3 | 0.15 | 0.077 | 29 | 29995 | 6.2 | 6 |
| 546 | Alpha-gliadin Bu-14 | 3 | 0.18 | 0.029 | 47 | 29995 | 6.2 | 6 |
| Total | 2.48 | |||||||
| Alpha-gliadins (Gli-B2) | ||||||||
| 327 | Alpha-gliadin Bu-11 | 5 | 1.59 | 0.107 | 30 | 34599 | 6.5 | 4 |
| 525 | Alpha-gliadin Bu-12 | 4 | 0.56 | 0.097 | 65 | 31541 | 7.0 | - |
| 328 | Alpha-gliadin Bu-12 | 4 | 0.27 | 0.058 | 25 | 31541 | 7.0 | - |
| 329 | Alpha-gliadin Bu-12 | 4 | 1.59 | 0.172 | 27 | 31541 | 7.0 | - |
| 387 | Alpha-gliadin Bu-12 | 4 | 0.17 | 0.017 | 49 | 31541 | 7.0 | - |
| 524 | Alpha-gliadin Bu-12 | 4 | 0.47 | 0.032 | 80 | 31541 | 7.0 | - |
| 341 | Alpha-gliadin Bu-23 | 3 | 3.29 | 0.154 | 59 | 33871 | 7.1 | - |
| Total | 7.94 | |||||||
| Alpha-gliadins (Gli-D2) | ||||||||
| 342 | Alpha-gliadin Bu-1 | 4 | 0.85 | 0.059 | 60 | 33412 | 7.8 | 1,2,3,4,5 |
| 330 | Alpha-gliadin Bu-210 | 6 | 2.66 | 0.148 | 52 | 30807 | 7.7 | 1,2,4 |
| 338 | Alpha-gliadin Bu-210 | 2 | 0.56 | 0.034 | 60 | 30808 | 7.7 | 1,2,4 |
| 468 | Alpha-gliadin Bu-3 | 4 | 1.66 | 0.119 | 71 | 33156 | 7.0 | 1,2,3,4,5 |
| 467 | Alpha-gliadin Bu-4 | 6 | 2.01 | 0.183 | 78 | 31462 | 6.6 | 1,2,3,4 |
| 550 | Alpha-gliadin Bu-10 | 4 | 1.31 | 0.127 | 75 | 31551 | 6.8 | 1,2,3 |
| Total | 9.05 | |||||||
| Alpha-gliadins (unknown locus) | ||||||||
| 124 | Alpha-gliadin Bu [GenBank:BQ807130] | 1 | 0.29 | 0.024 | inc | inc | inc | inc |
| 177 | Alpha-gliadin Bu [GenBank:BQ806209] | 5 | 0.32 | 0.118 | inc | inc | inc | inc |
| 420 | Alpha-gliadin Bu-27 | 3 | 0.34 | 0.055 | 55 | 34268 | 8.3 | - |
| Total | 0.96 | |||||||
| Gliadin mixed spots11 | ||||||||
| 334 | 3 gamma-gliadins, 1 alpha-gliadin | 4 | 1.08 | 0.105 | nd | nd | nd | nd |
| 389 | 1 alpha-gliadin, 1 other protein | 2 | 0.14 | 0.009 | nd | nd | nd | nd |
| 172 | 1 gamma-gliadin, 2 alpha-gliadins | 3 | 1.21 | 0.100 | nd | nd | nd | nd |
| 530 | 1 globulin, 1 alpha-gliadin | 2 | 0.08 | 0.019 | nd | nd | nd | nd |
| 195 | 1 farinin, 1 alpha-gliadin, 1 enzyme | 3 | 0.05 | 0.018 | nd | nd | nd | nd |
| Total | 2.56 |
Loci are indicated within parentheses.
1-7 Footnotes as in Table 3.
8inc indicates sequence was incomplete. nd indicates not determined because the spot contained a mixture of proteins.
9 The numbers indicate the presence of one or more of the following peptide sequences associated with celiac disease: 1, glia-α-9 epitope, core sequence PFPQPQLPY; 2, glia-α-20 epitope, core sequence PQPQLPYPQ; 3, glia-α-20 epitope, core sequence RPQQPYPQ; 4, glia-α epitope, core sequence QGSFQPSQQ; 5, 33-mer toxic fragment, LQLQPFPQPQLPYPQPQLPYPQPQLPYPQPQPF; 6, p31-43, LGQQQPFPPQQPY. - indicates that the alpha-gliadin does not contain any of these sequences.
10Contains seven Cys residues.
11No single protein had a predominant number of peptides. Details are in Additional file 1.