Phosphorylation of paxillin LD4 destabilizes helix formation and inhibits binding to focal adhesion kinase.
Cell migration is a dynamic process that requires the coordinated formation and disassembly of focal adhesions (FAs). Several proteins such as paxillin, focal adhesion kinase (FAK), and G protein-coupled receptor kinase-interacting protein 1 (GIT1) are known to play a regulatory role in FA disassembly and turnover. However, the mechanisms by which this occurs remain to be elucidated. Paxillin has been shown to bind the C-terminal domain of FAK in FAs, and an increasing number of studies have linked paxillin association with GIT1 during focal adhesion disassembly. It has been reported recently that phosphorylation of serine 273 in the LD4 motif of paxillin leads to an increased association with Git1 and focal adhesion turnover. In the present study, we examined the effects of phosphorylation of the LD4 peptide on its binding affinity to the C-terminal domain of FAK. We show that phosphorylation of LD4 results in a reduction of binding affinity to FAK. This reduction in binding affinity is not due to the introduction of electrostatic repulsion or steric effects but rather by a destabilization of the helical propensity of the LD4 motif. These results further our understanding of the focal adhesion turnover mechanism as well as identify a novel process by which phosphorylation can modulate intracellular signaling.
Bertolucci
Craig M
CM
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
Guibao
Cristina D
CD
Zheng
Jie J
JJ
D020816:Amino Acid Motifs; D000818:Animals; D016880:Anisotropy; D002645:Chickens; D002942:Circular Dichroism; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism*; D008958:Models, Molecular; D051419:Paxillin / Q000737:chemistry* / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D010766:Phosphorylation; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence
D051419:Paxillin; D010455:Peptides; D051416:Focal Adhesion Protein-Tyrosine Kinases