Defining the intramembrane binding mechanism of sarcolipin to calcium ATPase using solution NMR spectroscopy
Defining the intramembrane binding mechanism of sarcolipin to calcium ATPase using solution NMR spectroscopy
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DOI：
10.1016/j.jmb.2006.02.005
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发表时间：
2006-04-28
影响因子：
5.6
通讯作者：
Veglia, G
中科院分区：
文献类型：
--
作者：
Buffy, JJ;Buck-Koehntop, BA;Veglia, G
Sarcolipin（SIN）是一种在骨骼肌和心肌中表达的完整膜蛋白，在骨骼肌和心肌中，其通过降低其表观Ca 2+亲和力以类似于其同源物受磷蛋白（PLN）的方式抑制SERCA（钙ATP酶）。我们使用溶液NMR映射SLN内发生的结构变化后，与监管目标，SERCA，共重组的两种蛋白质在十二烷基磷酸胆碱（DPC）洗涤剂胶束，一个系统，保留了天然结构的SIN和SERCA的活性，与比较这些相互作用与先前研究的PLN-SERCA复合物的目标。我们的结构动力学分析的SLN在DPC胶束显示，这种多肽被分配成四个亚结构域：一个短的非结构化的N端（残基1-6），一个短的动态螺旋（残基7-14），一个更刚性的螺旋（残基15-26），和一个非结构化的C端（残基27-31）。在添加SERCA后，不同的结构域根据其动力学表现，在酶的表面上成型。值得注意的是，SIN的每个结构域以类似于PLN中相应结构域的方式表现，支持SIN和PLN在相同沟中以类似机制结合SERCA的假设。(c)2006爱思唯尔有限公司保留所有权利。
Sarcolipin (SIN) is an integral membrane protein that is expressed in both skeletal and cardiac muscle, where it inhibits SERCA (calcium ATPase) by lowering its apparent Ca2+ affinity in a manner similar to that of its homologue phospholamban (PLN). We use solution NMR to map the structural changes occurring within SLN upon interaction with the regulatory target, SERCA, co-reconstituting the two proteins in dodecylphosphocholine (DPC) detergent micelles, a system that preserves the native structure of SIN and the activity of SERCA, with the goal of comparing these interactions with those of the previously studied PLN-SERCA complex. Our analysis of the structural dynamics of SLN in DPC micelles shows this polypeptide to be partitioned into four subdomains: a short unstructured N terminus (residues 1-6), a short dynamic helix (residues 7-14), a more rigid helix (residues 15-26), and an unstructured C terminus (residues 27-31). Upon addition of SERCA, the different domains behave according to their dynamics, molding onto the surface of the enzyme. Remarkably, each domain of SIN behaves in a manner similar to that of the corresponding domains in PLN, supporting the hypothesis that both SIN and PLN bind SERCA in the same groove and with similar mechanisms. (c) 2006 Elsevier Ltd. All rights reserved.