Phosphorylation of the vasodilator-stimulated phosphoprotein regulates its interaction with actin

被引:205
作者
Harbeck, B [1 ]
Hüttelmaier, S [1 ]
Schlüter, K [1 ]
Jockusch, BM [1 ]
Illenberger, S [1 ]
机构
[1] Tech Univ Carolo Wilhelmina Braunschweig, Inst Zool, Dept Cell Biol, Bioctr, D-38092 Braunschweig, Germany
关键词
D O I
10.1074/jbc.M005066200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The vasodilator-stimulated phosphoprotein (VASP) is a major substrate for cyclic nucleotide-dependent kinases in platelets and other cardiovascular cells. It promotes actin nucleation and binds to actin filaments in vitro and associates with stress fibers in cells. The VASP-actin interaction is salt-sensitive, arguing for electrostatic interactions. Hence, phosphorylation may significantly alter the actin binding properties of VASP. This hypothesis was investigated by analyzing complex formation of recombinant murine VASP with actin after phosphorylation with cAMP-dependent kinase in different assays. cAMP-dependent kinase phosphorylation had a negative effect on both actin nucleation and VASP interaction with actin filaments, with the actin nucleating capacity being more affected than actin filament binding and bundling. Replacing VASP residues known to be phosphorylated in vivo by acidic residues to mimic phosphorylation had similar although less dramatic effects on VASP-actin interactions. In contrast, phosphorylation had no significant effect on VASP oligomerization or its interaction with its known ligands profilin, vinculin, and zyxin. When overexpressing VASP mutants in eukaryotic cells, they all showed targeting to focal contacts and stress fibers, Our results imply that VASP phosphorylation may act as an immediate negative regulator of actin dynamics.
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收藏
页码:30817 / 30825
页数:9
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