کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1925569 | 1536393 | 2012 | 9 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Cofilin weakly interacts with 14-3-3 and therefore can only indirectly participate in regulation of cell motility by small heat shock protein HspB6 (Hsp20) Cofilin weakly interacts with 14-3-3 and therefore can only indirectly participate in regulation of cell motility by small heat shock protein HspB6 (Hsp20)](/preview/png/1925569.png)
It has been previously reported that phosphorylated cofilin interacted with 14-3-3ζ protein to generate a sub-micromolar Kd binary complex. Here we challenge this hypothesis by analyzing the direct association of recombinant cofilin with 14-3-3ζ using different in vitro biochemical methods. Phosphorylated cofilin at high concentration binds to 14-3-3 immobilized on nitrocellulose, however no complex formation was detected by means of native gel electrophoresis or chemical crosslinking. Intact dimeric or mutant monomeric 14-3-3 was unable to form stable complexes with phosphorylated or unphosphorylated cofilin detected by size-exclusion chromatography. In co-sedimentation assay 14-3-3 did not affect interaction of cofilin with F-actin. The data of native gel electrophoresis indicate that 14-3-3 did not affect interaction of cofilin with G-actin. Thus, cofilin only weakly interacts with 14-3-3 and therefore cannot directly compete with phosphorylated small heat shock protein HspB6 for its binding to 14-3-3. It is hypothesized that phosphorylated HspB6 might affect interaction of 14-3-3 with protein phosphatases (and/or protein kinases) involved in dephosphorylation (or phosphorylation) of cofilin and by this means regulate cofilin-dependent reorganization of cytoskeleton.
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► Phosphorylated cofilin interacted with 14-3-3 bound to nitrocellulose.
► Native electrophoresis and crosslinking failed to detect cofilin-14-3-3 complex.
► Cofilin did not interact either with dimeric or monomeric 14-3-3.
► 14-3-3 did not affect interaction of cofilin with either G- or F-actin.
► Cofilin cannot compete with phosphorylated HspB6 for 14-3-3 binding.
Journal: Archives of Biochemistry and Biophysics - Volume 521, Issues 1–2, May 2012, Pages 62–70