Lu X, Xu L, Meissner G
Department of Biochemistry, University of North Carolina, Chapel Hill 27599-7260.
J Biol Chem. 1994 Mar 4;269(9):6511-6.
Expression studies with skeletal and cardiac muscle cDNAs have suggested that the putative cytoplasmic loop region of the dihydropyridine receptor (DHPR) alpha 1 subunit between transmembrane repeats II and III (DCL) is a major determinant of the type of excitation-contraction coupling (skeletal or cardiac) in rescued dysgenic muscle cells (Tanabe, T., Beam, K. G., Adams, B. A., Niidome, T., and Numa, S. (1990) Nature 346, 567-569). In this study, the possibility of a direct functional interaction with the sarcoplasmic reticulum ryanodine receptor/Ca2+ release channel has been tested by expressing the DCLs of the mammalian skeletal and cardiac muscle DHPR alpha 1 subunit in Escherichia coli. The purified peptides activated the skeletal muscle ryanodine receptor/Ca2+ release channel in single channel and [3H]ryanodine binding measurements, by increasing channel open probability and the affinity of [3H]ryanodine binding, respectively. The two peptides did not activate the cardiac muscle Ca2+ release channel. Other proteins (polylysine, serum albumin) also increased [3H]ryanodine binding and Ca2+ release channel activity, but their activation mechanisms were distinguishable from DCLs. These results show that the II-III cytoplasmic loop of the skeletal and cardiac DHPR alpha 1 subunit functionally interacts with the skeletal, but not cardiac, muscle Ca2+ release channel. Furthermore, our studies suggest that in addition to the DHPR, the sarcoplasmic reticulum Ca2+ release channel may determine the type of E-C coupling that exists in muscle.
对骨骼肌和心肌cDNA的表达研究表明,二氢吡啶受体(DHPR)α1亚基跨膜重复序列II和III之间的假定胞质环区域(DCL)是拯救的发育不全肌细胞中兴奋-收缩偶联类型(骨骼肌或心肌)的主要决定因素(田边,T.,比姆,K.G.,亚当斯,B.A.,新留,T.,和沼田,S.(1990)《自然》346,567 - 569)。在本研究中,通过在大肠杆菌中表达哺乳动物骨骼肌和心肌DHPR α1亚基的DCL,测试了其与肌浆网兰尼碱受体/Ca2 +释放通道直接功能相互作用的可能性。纯化的肽在单通道和[3H]兰尼碱结合测量中激活了骨骼肌兰尼碱受体/Ca2 +释放通道,分别通过增加通道开放概率和[3H]兰尼碱结合亲和力来实现。这两种肽未激活心肌Ca2 +释放通道。其他蛋白质(聚赖氨酸、血清白蛋白)也增加了[3H]兰尼碱结合和Ca2 +释放通道活性,但它们的激活机制与DCL不同。这些结果表明,骨骼肌和心肌DHPR α1亚基的II - III胞质环与骨骼肌而非心肌的Ca2 +释放通道存在功能相互作用。此外,我们的研究表明,除了DHPR外,肌浆网Ca2 +释放通道可能也决定了肌肉中存在的兴奋 - 收缩偶联类型。