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 alpha-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 alpha-actinin, but not with HSP27 or paxillin. Immunostaining with HSP20 and alpha-actinin antibodies demonstrated that HSP20 and alpha-actinin co-localized. The amount of HSP20 that immunoprecipitated with alpha -actinin was markedly diminished in muscles that were treated with the vasorelaxant forskolin. CONCLUSIONS: HSP20 is associated with both actin and alpha-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 alpha-actinin. These data suggest that phosphorylation of HSP20 may lead to relaxation of vascular smooth muscles through a dynamic association with cytoskeletal elements.
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 alpha-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 alpha-actinin, but not with HSP27 or paxillin. Immunostaining with HSP20 and alpha-actinin antibodies demonstrated that HSP20 and alpha-actinin co-localized. The amount of HSP20 that immunoprecipitated with alpha -actinin was markedly diminished in muscles that were treated with the vasorelaxant forskolin. CONCLUSIONS:HSP20 is associated with both actin and alpha-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 alpha-actinin. These data suggest that phosphorylation of HSP20 may lead to relaxation of vascular smooth muscles through a dynamic association with cytoskeletal elements.
Authors: Charles R Flynn; Christopher C Smoke; Elizabeth Furnish; Padmini Komalavilas; Jeffrey Thresher; Zhengping Yi; Lawrence J Mandarino; Colleen M Brophy Journal: Protein Expr Purif Date: 2006-09-12 Impact factor: 1.650
Authors: Alex Banathy; Joyce Cheung-Flynn; Kasia Goleniewska; Kelly L Boyd; Dawn C Newcomb; R Stokes Peebles; Padmini Komalavilas Journal: Am J Respir Cell Mol Biol Date: 2016-08 Impact factor: 6.914
Authors: Padmini Komalavilas; Raymond B Penn; Charles R Flynn; Jeffrey Thresher; Luciana B Lopes; Elizabeth J Furnish; Manhong Guo; Manuel A Pallero; Joanne E Murphy-Ullrich; Colleen M Brophy Journal: Am J Physiol Lung Cell Mol Physiol Date: 2007-11-09 Impact factor: 5.464