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Table 1.

Phosphorylation sites of murine βPIXa detected by mass spectrometry

Site Detected in serum-incubated sample Detected in EGF-stimulated sample Homologs/other family members Putative kinases
S71   +   +   R, M, H/αPIX   CDC2, CDK5  
S79   Not covered   +   R, M, H   CDC2, CDK5  
S131   +   +   R, M, H   
S135   +    R, M, H   
S145   +    R, M   
S146   +    R, M, H   
S192   +   +   R, M, H/αPIX   
S195   +   +   R, M   
S340    +   R, M, H/αPIX   
S427/S428    +   R, M, H/αPIX (S428 only)   
S498   IMAC   Not covered   R, M, H/αPIX   CDC2, CDK5  
S516   +   +   R, M, H/αPIX   PKA  
Y542   IMAC    R, M, H/αPIX   
S559   IMAC   IMAC   R, M, H/αPIX   PKA, AKT, 14-3-3  
S588   Not covered   +   R, M, H/αPIX   PKCmu  
T592    +   R, M, H/αPIX   
Site Detected in serum-incubated sample Detected in EGF-stimulated sample Homologs/other family members Putative kinases
S71   +   +   R, M, H/αPIX   CDC2, CDK5  
S79   Not covered   +   R, M, H   CDC2, CDK5  
S131   +   +   R, M, H   
S135   +    R, M, H   
S145   +    R, M   
S146   +    R, M, H   
S192   +   +   R, M, H/αPIX   
S195   +   +   R, M   
S340    +   R, M, H/αPIX   
S427/S428    +   R, M, H/αPIX (S428 only)   
S498   IMAC   Not covered   R, M, H/αPIX   CDC2, CDK5  
S516   +   +   R, M, H/αPIX   PKA  
Y542   IMAC    R, M, H/αPIX   
S559   IMAC   IMAC   R, M, H/αPIX   PKA, AKT, 14-3-3  
S588   Not covered   +   R, M, H/αPIX   PKCmu  
T592    +   R, M, H/αPIX   

Serum-incubated and EGF-stimulated samples were both treated with calyculin A and peroxovanadate to inhibit phosphatases. Phosphorylation sites labeled `IMAC' were only detected after sample enrichment by immobilized metal affinity chromatography (IMAC). Sites detected without enrichment are denoted with a `+'. `Not covered' signifies that the residue was not observed during mass spectrometry analysis, therefore phosphorylation status could not be determined. Homologous phosphorylation sites were determined via sequence comparison analysis using FASTA (http://fasta.bioch.virginia.edu/) (Pearson, 1990). Consensus phosphorylation motifs for putative kinases were analyzed using the Scansite scoring algorithm (http://scansite.mit.edu) (Obenauer et al., 2003). For additional data regarding βPIXa phosphorylation, see the Cell Migration Consortium website: http://www.cellmigration.org. R, rat; M, mouse; H, human.

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