Broad disorder and the allosteric mechanism of myosin II regulation by phosphorylation.
Broad disorder and the allosteric mechanism of myosin II regulation by phosphorylation.
复制标题
肌球蛋白 II 磷酸化调节的广泛紊乱和变构机制。
DOI：
10.1073/pnas.1014137108
复制
发表时间：
2011
影响因子：
11.1
通讯作者：
Fajer,PiotrG
中科院分区：
文献类型：
--
作者：
Vileno,Bertrand;Chamoun,Jean;Liang,Hua;Brewer,Paul;Haldeman,BrianD;Facemyer,KevinC;Salzameda,Bridget;Song,Likai;Li,Hui-Chun;Cremo,ChristineR;Fajer,PiotrG
Double electron electron resonance EPR methods was used to measure the effects of the allosteric modulators, phosphorylation, and ATP, on the distances and distance distributions between the two regulatory light chain of myosin (RLC). Three different states of smooth muscle myosin (SMM) were studied: monomers, the short-tailed subfragment heavy meromyosin, and SMM filaments. We reconstituted myosin with nine single cysteine spin-labeled RLC. For all mutants we found a broad distribution of distances that could not be explained by spin-label rotamer diversity. For SMM and heavy meromyosin, several sites showed two heterogeneous populations in the unphosphorylated samples, whereas only one was observed after phosphorylation. The data were consistent with the presence of two coexisting heterogeneous populations of structures in the unphosphorylated samples. The two populations were attributed to an on and off state by comparing data from unphosphorylated and phosphorylated samples. Models of these two states were generated using a rigid body docking approach derived from EM [Wendt T, Taylor D, Trybus KM, Taylor K (2001)Proc Natl Acad Sci USA98:4361–4366] (PNAS, 2001, 98:4361–4366), but our data revealed a new feature of the off-state, which is heterogeneity in the orientation of the two RLC. Our average off-state structure was very similar to the Wendt model reveal a new feature of the off state, which is heterogeneity in the orientations of the two RLC. As found previously in the EM study, our on-state structure was completely different from the off-state structure. The heads are splayed out and there is even more heterogeneity in the orientations of the two RLC.
