Table 1. Mass spectrometric identifications from all three AP-1γ IPs.
Protein | Experiment 1 | Experiment 2 | Experiment 3 | |||||||||
Score | Peptides | Score | Peptides | Score | Peptides | |||||||
Soft | Std | Soft | Std | Soft | Std | Soft | Std | Soft | Std | Soft | Std | |
AP-1γ1 | 726 | 635 | 28 | 37 | 2757 | 1737 | 118 | 77 | 2582 | 3450 | 86 | 147 |
AP-1β | 2072 | 1779 | 92 | 109 | 3792 | 2655 | 160 | 111 | 5854 | 4996 | 244 | 212 |
AP-1µ1a | 495 | 1348 | 44 | 96 | 2906 | 1754 | 148 | 100 | 3124 | 3650 | 145 | 178 |
AP-1σ1a | 901 | 1406 | 14 | 26 | 839 | 383 | 25 | 11 | 777 | 978 | 24 | 24 |
AP-1σ1b | 74 | 62 | 5 | 6 | 141 | 68 | 9 | 3 | 162 | 224 | 15 | 11 |
AP-1σ1c | 98 | - | 2 | - | 31 | - | 1 | - | 141 | 91 | 11 | 3 |
Aftiphilin | 332 | 241 | 20 | 9 | 1235 | 519 | 34 | 20 | 2622 | 2644 | 88 | 83 |
γ-Synergin | 1052 | 549 | 21 | 18 | 1068 | 404 | 29 | 16 | 5596 | 3881 | 188 | 138 |
p200 | 282 | 354 | 13 | 12 | 903 | 494 | 39 | 17 | 4116 | 4074 | 161 | 165 |
KIF13a | 745 | 524 | 29 | 21 | 1386 | 851 | 59 | 30 | 280 | 191 | 12 | 11 |
p34 | 63 | 50 | 2 | 2 | 330 | 269 | 19 | 14 | 105 | 83 | 3 | 3 |
This table shows AP-1 constituents and associated proteins identified in three independent AP1-γ immunoprecipitations. In each experiment, samples (ie Protein A sepharose beads with bound antigen) were split into two equal aliquots prior to elution. Aliquots were either subjected to the soft elution protocol (“Soft”) reported in this study, or to standard elution conditions (“Std”). Eluates were subjected to tryptic solution digest without further fractionation, and analyzed by LC-MSMS. Mascot scores reflecting the confidence of identification [8], and the number of peptides identified for each protein are indicated. Please note that the AP-1σ1c subunit was not identified in experiments 1 and 2 using standard elution conditions.
Experiments 1 and 2 were performed in MES-D buffer; experiment 3 was performed in PBS-T buffer (see File S1 for details).