The small heat shock protein (HSP) 20 is dynamically associated with the actin cross-linking protein actinin

Abstract

Background. The heat shock-related protein (HSP) 20 is associated with actin and modulates smooth-muscle relaxation. We hypothesized that HSP20 mediates vasorelaxation via dynamic interactions with cytoskeletal proteins, such as actin, or actin binding proteins, such as α-actinin. Methods. Physiological responses of strips of bovine carotid artery were analyzed with a muscle bath. In other experiments, the arteries were homogenized, and imunoprecipitations were performed. Immunohistochemistry with anti-HSP20 and anti-actinin antibodies was used to determine co-localization of the two proteins. Results. Bovine carotid arteries contracted in response to serotonin and rapidly relaxed in response to forskolin. HSP20 co-immunoprecipitated with both actin and α-actinin, but not with HSP27 or paxillin. Immunostaining with HSP20 and α-actinin antibodies demonstrated that HSP20 and α-actinin co-localized. The amount of HSP20 that immunoprecipitated with α -actinin was markedly diminished in muscles that were treated with the vasorelaxant forskolin. Conclusions. HSP20 is associated with both actin and α-actinin. Activation of cyclic nucleotide-dependent signaling pathways leads to increases in the phosphorylation of HSP20 and a decrease in the association of HSP20 with α-actinin. These data suggest that phosphorylation of HSP20 may lead to relaxation of vascular smooth muscles through a dynamic association with cytoskeletal elements.