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首页> 外文期刊>American Journal of Physiology >Differences in calmodulin and calmodulin-binding proteins in phasic and tonic smooth muscles.
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Differences in calmodulin and calmodulin-binding proteins in phasic and tonic smooth muscles.

机译：钙调蛋白和钙调蛋白结合蛋白在阶段性和张力性平滑肌中的差异。

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摘要

To determine whether densities of calmodulin (CaM) and CaM-binding proteins are related to phasic and tonic behavior of smooth muscles, we quantified these proteins in the opossum esophageal body (EB) and lower esophageal sphincter (LES), which represent phasic and tonic smooth muscles, respectively. Gel electrophoresis, immunoprecipitation, Western blot, and hemagglutinin epitope-tagged CaM (HA-CaM) overlay assay with quantitative scanning densitometry and phosphorylation measurements were used. Total protein content in the two smooth muscles was similar (approximately 30 mg protein/g frozen tissue). Total tissue concentration of CaM was significantly (25%) higher in EB than in LES (P < 0.05). HA-CaM-binding proteins were qualitatively similar in LES and EB extracts. Myosin, myristoylated alanine-rich C kinase substrate protein, Ca(2+)/CaM kinase II, and calponin contents were also similar in the two muscles. However, content and total activity of myosin light chain kinase (MLCK) and content of caldesmon (CaD) were three- to fourfold higher in EB than in LES. Increased CaM and MLCK content may allow for a wide range of contractile force varying from complete relaxation in the basal state to a large-amplitude, high-velocity contraction in EB phasic muscle. Increased content of CaD, which provides a braking mechanism on contraction, may further contribute to the phasic contractile behavior. In contrast, low CaM, MLCK, and CaD content may be responsible for a small range of contractile force seen in tonic muscle of LES.
机译：为了确定钙调蛋白（CaM）和CaM结合蛋白的密度是否与平滑肌的相变和张力行为有关，我们在负鼠食管体（EB）和下食道括约肌（LES）中定量了这些蛋白，代表了相变和张力分别是平滑肌。使用凝胶电泳，免疫沉淀，Western印迹和血凝素表位标记的CaM（HA-CaM）重叠测定，并进行定量扫描光密度法和磷酸化测量。两条平滑肌中的总蛋白含量相似（大约30 mg蛋白/ g冷冻组织）。 EB中CaM的总组织浓度显着（25％）高于LES（P <0.05）。 LES和EB提取物中的HA-CaM结合蛋白在质量上相似。肌球蛋白，肉豆蔻酰化的富含丙氨酸的C激酶底物蛋白，Ca（2 +）/ CaM激酶II和钙蛋白的含量在两条肌肉中也相似。但是，EB中肌球蛋白轻链激酶（MLCK）的含量和总活性以及caldesmon（CaD）的含量比LES高三到四倍。 CaM和MLCK含量的增加可能允许范围广泛的收缩力，从基础状态的完全松弛到EB相肌的大幅度，高速收缩。 CaD含量的增加，为收缩提供了一种制动机制，可能进一步有助于该阶段的收缩行为。相反，低水平的CaM，MLCK和CaD含量可能是导致LES的强直性肌肉中出现小范围收缩力的原因。

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