Table 1.

Summary of paxillin phosphorylation sites

FLAG-GFP-Pax lower-level expression (LTQ-FT) Detected without inhibitors and without enrichment (trypsin) Homologs/other family members Putative kinases
Y31 mpax and hpax Src/FAK§
Y40 mpax and hpax Src
GlcNAc S74 Yes (ETD) mpax and hpax
S83 mpax and hpax
S85 Yes hpax p38MAPK**, GSK3
Y88 mpax and hpax Src
S89 mpax
S96 mpax and hpax
S98 mpax and hpax PKC
S106 Yes mpax and hpax ERK, GSK3
S108 Yes hic-5 PKC, GSK3
S112 - PKA
Y118 mpax, hpax and hic-5 Src/FAK††,§
S119 mpax, hpax and hic-5
S126 Yes mpax, hpax and hic-5 GSK3
S130 Yes mpax and hpax
S137 mpax, hpax and hic-5
S164
S171 Yes mpax, hpax and hic-5
S173 Yes PKA, Akt, CaMII
Y182 mpax and hpax EGFR, PDFGR
S187-190
S227/228 mpax and hpax GSK3 (S227)
S231 mpax, hpax and hic-5 ERK
S239
S245 mpax and hpax p34cdc2, Cdk5
S259 mpax, hpax and hic-5 PKA
S290 Yes mpax, hpax and hic-5 ERK
S382 mpax, hpax and hic-5 PKC
S501 Yes mpax, hpax and hic-5 PKA, CaMII
Additional sites detected
FLAG-Pax overexpression (LCQ-XP)* Comments (multiple enzymes) Homologs/other family members Putative kinases
T29 Observed once hpax
S94 GSK3
S143 Observed once mpax and hpax
T199 hpax
S262 mpax, hpax and hic-5
S273 mpax, hpax and hic-5
T295 hic-5 PKC, Cdk5
S308 mpax and hpax
FLAG-Pax overexpression (LTQ-FT or ETD)* Comments (trypsin) Homologs/other family members Putative kinases
Y76 mpax and hic-5
S90 ETD mpax and hpax GSK3
S91 ETD mpax and hpax PKC
S217/220 mpax and hpax PKA (S217)
S275 mpax, hpax and hic-5
T344 ETD mpax and hpax
S361 ETD mpax and hpax
Y377 mpax, hpax and hic-5
T403 ETD mpax and hpax
Y436 mpax and hpax
S504 ETD mpax and hpax
1540 ETD mpax and hpax PKC
  • The table shows phosphorylation sites detected in chicken paxillin by mass spectrometry (the numbering is according to the chicken paxillin sequence).

  • * The FLAG-paxillin sequence (the FLAG sequence is not included): MDDLDALLADLESTTSHISKRPVFLTEETPYSYPTGNHTYQEIAVPPPVPPPPSSEALNGTDPLDQWQPSVSRYGHQ QPQSQSPIYSSSAKSSSASVPRDGLSSPSPRASEEEHVYSFPNKQKSAEPSPTMTSTSLGSNLSELDRLLLELNAVQ HNPPSGFSADEVSRSPSLPNVTGPHYVIPESSSSAGGKAAPPTKEKPKRNGGRGIEDVRPSVESLLDELESSVPSPV PAITVSQGEVSSPQRVNASQQQTRISASSATRELDELMASLSDFKFMAQGKAGGSSSPPSTTPKPGSQLDTMLGSLQ SDLNKLGVATVAKGVCGACKKPIAGQVVTAMGKTWHPEHFVCTHCQEEIGSRNFFERDWQPYCEKYYHNLFSPRCYY CNGPILDKVVTAFDRTWHPEHFFCAQCGVFFGPEGFHEKDGKAYCRKDYFDMFAPKCGGCARAILENYISALNTLWH PECFVCRECFTPFINGSFFEHDGQPYCEVHYHERRGSLCSGCQKPITGRYITAMGKKFHPEHFVCAFCLKQLNKGTF KEQNYKPYCQNCFLKLFC. Using multiple enzymes, 97% coverage of this sequence was detected by mass spectrometry

  • This phosphopeptide contained an amino acid that differed from the corresponding FLAG-GFP peptide sequence

  • This column shows the kinases predicted to phosphorylate the indicated sites using NetPhos 2.0 and Scansite. Note that this is a partial list; we have only included potential kinases that have been implicated in cell migration. References are given for kinases that have been shown experimentally to phosphorylate the site

    LCQ-XP, dynamic range approximately 100, collisional-activated dissociation MS/MS spectra. LTQ-FT, dynamic range approximately 5000, collisional activated dissociation MS/MS spectra. ETD, electron transfer dissociation. Peptide fragmentation by ETD enabled phosphorylation or glycosylation site identification.

    For additional data regarding paxillin phosphorylation, see the Cell Migration Consortium web site (http://www.cellmigration.org).

  • § Schaller and Parsons, 1995

  • Schaller and Schaefer, 2001

  • ** Huang, et al., 2004

  • †† Bellis et al., 1995