Transduction of biologically active motifs of the small heat shock-related protein, HSP20, leads to relaxation of vascular smooth muscle
Transduction of biologically active motifs of the small heat shock-related protein, HSP20, leads to relaxation of vascular smooth muscle
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DOI：
10.1096/fj.02-1028fje
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发表时间：
2003-05-01
期刊：
影响因子：
4.8
通讯作者：
Brophy, CM
中科院分区：
文献类型：
--
作者：
Flynn, CR;Komalavilas, P;Brophy, CM
环核苷酸依赖性信号通路的激活导致热休克相关小蛋白 HSP20 在丝氨酸 16 上磷酸化，并松弛血管平滑肌。在这项研究中，我们使用增强的蛋白转导结构域 (PTD) 序列将 HSP20 磷酸肽类似物递送到猪冠状动脉中。磷酸HSP20类似物的转导导致冠状动脉平滑肌的剂量依赖性松弛。含有与随机方向的相同氨基酸偶联的蛋白质转导结构域的肽则不然。与异硫氰酸荧光素 (FITC)-PTD 或 FITC-PTD-HSP20 肽一起孵育的动脉环的直接荧光显微镜显示显示弥漫性肽摄取。使用平滑肌匀浆的质谱免疫分析 (MSIAs) 来确定磷酸肽类似物是否影响内源性 HSP20 的磷酸化。用磷酸二酯酶抑制剂罂粟碱处理导致质量转移 80 Da。然而，在用磷酸化 HSP20 类似物处理的肌肉中，HSP20 的质量没有变化。这表明单独的PTD-磷酸化HSP20肽足以抑制力维持，并且可能对磷酸化HSP20的靶标具有直接作用。这些结果表明 HSP20 磷酸肽类似物的转导直接改变完整肌肉的生理反应。该数据还支持磷酸化 HSP20 在介导血管舒张中的直接作用。
Activation of cyclic nucleotide-dependent signaling pathways leads to phosphorylation of the small heat shock-related protein, HSP20, on serine 16, and relaxation of vascular smooth muscle. In this study, we used an enhanced protein transduction domain (PTD) sequence to deliver HSP20 phosphopeptide analogs into porcine coronary artery. The transduction of phosphoHSP20 analogs led to dose-dependent relaxation of coronary artery smooth muscle. Peptides containing the protein transduction domain coupled to a random orientation of the same amino acids did not. Direct fluorescence microscopy of arterial rings incubated with fluorescein isothiocyanate (FITC)-PTD or FITC-PTD-HSP20 peptides showed a diffuse peptide uptake. Mass spectrometric immunoassays (MSIAs) of smooth muscle homogenates were used to determine whether the phosphopeptide analogs affected the phosphorylation of endogenous HSP20. Treatment with the phosphodiesterase inhibitor papaverine led to a mass shift of 80 Da. However, there was no mass shift of HSP20 in muscles treated with phosphoHSP20 analogs. This suggests that the PTD-phosphoHSP20 peptide alone is sufficient to inhibit force maintenance and likely has a direct effect on the target of phosphorylated HSP20. These results suggest that transduction of phosphopeptide analogs of HSP20 directly alters physiological responses of intact muscles. The data also support a direct role for phosphorylated HSP20 in mediating vasorelaxation.