Table 1.
A meta-map of Gab2 phosphorylation
| Residue | This study | IM-treated K562 cells [[38]] | EGF-stimulated MCF-10A cells [[19]] | Phospho-sitePlus | Protein docking site or other known function | Conserved in Gab1 |
|---|---|---|---|---|---|---|
|
S2 |
X |
|
|
|
|
|
| Y48 |
|
X |
|
X |
|
X |
| Y49 |
|
|
|
X |
|
X |
|
S132 |
X |
|
|
|
|
|
| S133 |
X |
|
X |
X |
|
|
| S140 |
X |
|
X |
X |
|
|
| S141 |
X |
|
X |
X |
|
|
|
S146 |
X |
|
|
|
|
|
| S148 |
X |
|
X |
X |
|
|
| S149 |
X |
|
X |
X |
|
|
| S159 |
Western (Figure 1A) |
|
X |
X |
Negative regulatory site; AKT substrate [21] |
|
| S164 |
|
|
X |
X |
|
|
| T171 |
|
|
|
X |
|
X |
| Y194 |
|
|
|
X |
|
|
| S210 |
X |
|
X |
X |
14-3-3 binding and negative regulatory site [19,23] |
|
| S218 |
X |
|
X |
X |
|
X |
| S223 |
X |
|
X |
X |
|
|
|
T225 |
X |
|
|
|
|
|
| Y249 |
|
|
|
X |
CrkL/PLCγ [13] |
X |
| S250 |
|
|
|
X |
|
X |
| S264 |
X |
|
X |
X |
|
X and phosphorylated [57] |
| T265 |
|
|
|
X |
|
X |
| Y266 |
|
X |
|
X |
CrkL/PLCγ [13] |
X |
| T278 |
X |
|
X |
|
|
|
| S281 |
X |
|
X |
X |
|
|
|
T283 |
X |
|
|
|
|
|
|
S285 |
X |
|
|
|
|
|
| T287 |
X |
|
X |
X |
|
|
| Y293 |
|
X |
X |
X |
CrkL [13] |
X |
| T294 |
|
|
|
X |
|
|
|
T319 |
X |
|
|
|
|
X |
|
Y324 |
X |
|
|
|
CrkL/PLCγ [13] |
X |
|
T329 |
X |
|
|
|
|
X |
| T331 |
|
|
X |
|
|
X |
|
T343 |
X |
|
|
|
|
|
|
S368 |
X |
|
|
|
|
X |
| T385 |
X |
|
X |
X |
|
|
| T391 |
X |
|
X |
X |
14-3-3 binding and negative regulatory site [19,23] |
X and phosphorylated [57] |
| S404 |
X |
|
|
X |
|
X |
| S405 |
X |
|
X |
X |
|
X |
| T408 |
|
|
|
X |
|
|
| Y409 |
|
X |
|
X |
|
X |
| Y411 |
|
|
|
X |
|
|
|
S422 |
X |
|
|
|
|
X and phosphorylated [57] |
|
S425 |
X |
|
|
|
|
|
|
S427 |
X |
|
|
|
|
|
|
S443 |
X |
|
|
|
|
|
|
S448 |
X |
|
|
|
|
|
| Y452 |
X |
|
|
X |
p85 recruitment site; Resilient against IM and DST in K562 cells [9] |
X |
| S472 |
|
|
|
|
X |
X |
| S474 |
|
|
|
|
X |
X |
| Y476 |
|
|
|
|
X |
X |
| S480 |
X |
|
X |
X |
|
X |
| S488 |
X |
|
|
X |
|
X |
| Y491 |
X |
|
|
X |
|
|
|
S506 |
X |
|
|
|
|
|
| S543 |
X |
|
X |
X |
ERK dependent phosphorylation of the S543 equivalent critical for PH-domain mediated membrane recruitment of Gab1 [56]. ERK consensus motif (PxxSP) conserved in Gab2 |
X |
| S550 |
|
|
|
X |
|
|
| Y563 |
|
|
|
X |
|
|
| Y584 |
|
|
|
X |
p85 [13] |
X |
| S611 |
|
|
|
X |
|
|
| Y614 |
|
|
|
X |
Shp2 [13] |
X |
| S622 |
X |
|
X |
X |
|
X |
| S623 |
X |
|
X |
X |
|
|
| S625 |
X |
|
|
X |
|
X |
| S631 |
X |
|
|
X |
|
|
| T632 |
|
|
|
X |
|
|
| S633 |
|
|
|
X |
|
|
|
S634 |
X |
|
|
|
|
|
| T636 |
X |
|
|
X |
|
X |
| S637 |
X |
|
|
X |
|
|
| Y643 | X | X | Shp2 [13] | X |
Comparison of the phospho-maps of Gab2 from this study with those established for IM treated K562 cells [38], EGF stimulated MCF-10A cells [19] and the sites listed in the curated phosphosite database (http://www.phosphosite.org). Note that in addition to other entries, the entries from the MCF-10A cells are also represented in this database. See Wöhrle et al. (2009) for further details on the indicated docking sites. Gab2 phoshorylation sites that have a homotopic counterpart in Gab1 (incl. conserved S⇔T conversions) were identified by comparing the human Gab2 (NP_53679) and Gab1 (NP_997006.1) sequences using the BLASTP 2.2.27+ tool. However, it should be noted that in many cases only the phosphorylation site itself and not its context were conserved between both paralogues, like it was described for the T391/T387 in Gab2/Gab1 [19,57]. All new Gab2 phosphorylation sites, identified in this study were marked in bold.