Analysis of the Retention Times of CDB3 Peptides Generated by Endoproteinase Asp-N Digestion Using the Computer Program Peptidesort
| Position of Peptide in CDB3 Sequence . | Inclusive Residues . | Peptide Mr . | Predicted Retention Time at pH 2.1 . | Predicted Retention Time at pH 7.4 . | Net Charge . | Aro . | Acid . | Base . | Sulf . | Phil . | Phob . |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 4 | 10-22 | 1,688.8 | −19.5 | −90.7 | −7.0 | 1 | 7 | 0 | 2 | 9 | 4 |
| 7 | 38-44 | 707.7 | −6.0 | −16.0 | −2.0 | 0 | 2 | 0 | 0 | 5 | 2 |
| 2 | 6-6 | 133.1 | −2.8 | −8.2 | −1.0 | 0 | 1 | 0 | 0 | 1 | 0 |
| 24 | 369-369 | 133.1 | −2.8 | −8.2 | −1.0 | 0 | 1 | 0 | 0 | 1 | 0 |
| 3 | 7-9 | 425.4 | −2.1 | −19.0 | −2.0 | 1 | 2 | 0 | 0 | 2 | 1 |
| 1 | 1-5 | 648.7 | −0.4 | −25.0 | −2.0 | 0 | 2 | 0 | 1 | 3 | 2 |
| 6 | 25-37 | 1,453.5 | 4.0 | −34.1 | −4.0 | 0 | 4 | 0 | 1 | 9 | 4 |
| 5 | 23-24 | 230.2 | 5.2 | −2.1 | −1.0 | 0 | 1 | 0 | 0 | 2 | 0 |
| 12 | 143-148 | 755.8 | 7.5 | 3.0 | 0.0 | 0 | 1 | 1 | 0 | 5 | 1 |
| 16 | 223-234 | 1,230.4 | 8.9 | 3.6 | −1.0 | 0 | 2 | 1 | 0 | 5 | 7 |
| 23 | 363-368 | 571.6 | 12.6 | 5.9 | −1.0 | 0 | 1 | 0 | 0 | 3 | 3 |
| 22 | 355-362 | 916.0 | 18.2 | 22.4 | 0.0 | 2 | 1 | 1 | 0 | 4 | 4 |
| 10 | 122-136 | 1,571.7 | 23.5 | 7.3 | −2.0 | 0 | 2 | 0 | 0 | 7 | 8 |
| 11 | 137-142 | 825.9 | 24.8 | 16.0 | −1.0 | 2 | 2 | 1 | 0 | 3 | 3 |
| 15 | 205-222 | 1,790.9 | 29.5 | 14.6 | −2.0 | 0 | 3 | 1 | 0 | 11 | 7 |
| 20 | 316-324 | 944.1 | 32.1 | 21.4 | −1.0 | 0 | 1 | 0 | 1 | 5 | 4 |
| 25 | 370-379 | 1,146.3 | 32.6 | 17.0 | −1.0 | 1 | 1 | 0 | 0 | 6 | 4 |
| 13 | 149-182 | 3,637.2 | 35.2 | 8.8 | 0.0 | 0 | 5 | 5 | 0 | 16 | 18 |
| 21 | 325-354 | 3,405.9 | 35.2 | 33.1 | 2.0 | 1 | 3 | 5 | 0 | 19 | 11 |
| 18 | 277-296 | 2,298.6 | 36.5 | 38.9 | 1.0 | 2 | 2 | 3 | 1 | 9 | 11 |
| 19 | 297-315 | 2,137.4 | 36.6 | 13.2 | −2.0 | 2 | 3 | 1 | 1 | 8 | 11 |
| 14 | 183-204 | 2,454.7 | 38.6 | 1.5 | −3.0 | 1 | 3 | 0 | 1 | 15 | 7 |
| 8 | 45-66 | 2,584.9 | 40.6 | 0.9 | −2.0 | 2 | 3 | 1 | 1 | 13 | 9 |
| 9 | 67-121 | 6,562.4 | 127.6 | 63.7 | −1.0 | 6 | 8 | 7 | 1 | 28 | 27 |
| 17 | 235-276 | 4,713.5 | 153.3 | 75.9 | −5.0 | 4 | 7 | 2 | 0 | 17 | 25 |
| Position of Peptide in CDB3 Sequence . | Inclusive Residues . | Peptide Mr . | Predicted Retention Time at pH 2.1 . | Predicted Retention Time at pH 7.4 . | Net Charge . | Aro . | Acid . | Base . | Sulf . | Phil . | Phob . |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 4 | 10-22 | 1,688.8 | −19.5 | −90.7 | −7.0 | 1 | 7 | 0 | 2 | 9 | 4 |
| 7 | 38-44 | 707.7 | −6.0 | −16.0 | −2.0 | 0 | 2 | 0 | 0 | 5 | 2 |
| 2 | 6-6 | 133.1 | −2.8 | −8.2 | −1.0 | 0 | 1 | 0 | 0 | 1 | 0 |
| 24 | 369-369 | 133.1 | −2.8 | −8.2 | −1.0 | 0 | 1 | 0 | 0 | 1 | 0 |
| 3 | 7-9 | 425.4 | −2.1 | −19.0 | −2.0 | 1 | 2 | 0 | 0 | 2 | 1 |
| 1 | 1-5 | 648.7 | −0.4 | −25.0 | −2.0 | 0 | 2 | 0 | 1 | 3 | 2 |
| 6 | 25-37 | 1,453.5 | 4.0 | −34.1 | −4.0 | 0 | 4 | 0 | 1 | 9 | 4 |
| 5 | 23-24 | 230.2 | 5.2 | −2.1 | −1.0 | 0 | 1 | 0 | 0 | 2 | 0 |
| 12 | 143-148 | 755.8 | 7.5 | 3.0 | 0.0 | 0 | 1 | 1 | 0 | 5 | 1 |
| 16 | 223-234 | 1,230.4 | 8.9 | 3.6 | −1.0 | 0 | 2 | 1 | 0 | 5 | 7 |
| 23 | 363-368 | 571.6 | 12.6 | 5.9 | −1.0 | 0 | 1 | 0 | 0 | 3 | 3 |
| 22 | 355-362 | 916.0 | 18.2 | 22.4 | 0.0 | 2 | 1 | 1 | 0 | 4 | 4 |
| 10 | 122-136 | 1,571.7 | 23.5 | 7.3 | −2.0 | 0 | 2 | 0 | 0 | 7 | 8 |
| 11 | 137-142 | 825.9 | 24.8 | 16.0 | −1.0 | 2 | 2 | 1 | 0 | 3 | 3 |
| 15 | 205-222 | 1,790.9 | 29.5 | 14.6 | −2.0 | 0 | 3 | 1 | 0 | 11 | 7 |
| 20 | 316-324 | 944.1 | 32.1 | 21.4 | −1.0 | 0 | 1 | 0 | 1 | 5 | 4 |
| 25 | 370-379 | 1,146.3 | 32.6 | 17.0 | −1.0 | 1 | 1 | 0 | 0 | 6 | 4 |
| 13 | 149-182 | 3,637.2 | 35.2 | 8.8 | 0.0 | 0 | 5 | 5 | 0 | 16 | 18 |
| 21 | 325-354 | 3,405.9 | 35.2 | 33.1 | 2.0 | 1 | 3 | 5 | 0 | 19 | 11 |
| 18 | 277-296 | 2,298.6 | 36.5 | 38.9 | 1.0 | 2 | 2 | 3 | 1 | 9 | 11 |
| 19 | 297-315 | 2,137.4 | 36.6 | 13.2 | −2.0 | 2 | 3 | 1 | 1 | 8 | 11 |
| 14 | 183-204 | 2,454.7 | 38.6 | 1.5 | −3.0 | 1 | 3 | 0 | 1 | 15 | 7 |
| 8 | 45-66 | 2,584.9 | 40.6 | 0.9 | −2.0 | 2 | 3 | 1 | 1 | 13 | 9 |
| 9 | 67-121 | 6,562.4 | 127.6 | 63.7 | −1.0 | 6 | 8 | 7 | 1 | 28 | 27 |
| 17 | 235-276 | 4,713.5 | 153.3 | 75.9 | −5.0 | 4 | 7 | 2 | 0 | 17 | 25 |
Peptides are listed in their predicted order of elution from a reverse-phase HPLC column at pH 2.1. Also included are the positions of each peptide in the sequence of CDB3, the molecular weight of each peptide, their predicted relative retention times at pH 2.1 and 7.4, the net charge of each peptide, and the number of aromatic, acidic, basic, sulfur-containing, hydrophilic, and hydrophobic residues in each peptide.