Irving T, Bhattacharya S, Tesic I, Moore J, Farman G, Simcox A, Vigoreaux J, Maughan D
Department of Biological, Chemical and Physical Sciences, Illinois Institute of Technology, Chicago 60616, USA.
J Muscle Res Cell Motil. 2001;22(8):675-83. doi: 10.1023/a:1016336024366.
The similarity of amino acid sequence and motifs of the N-terminal extensions of certain class II myosin light chains, found throughout the animal kingdom, suggest a common functional role. One possible role of the N-terminal extension is to enhance oscillatory work and power production in striated muscles that normally operate in an oscillatory mode. We conducted small-angle X-ray diffraction experiments and small-length-perturbation analysis to examine the structural and functional consequences of deleting the N-terminal extension of the myosin regulatory light chain (RLC) in Drosophila flight muscle. The in vivo lattice spacing of dorsal longitudinal muscle (DLM) of flies lacking the RLC N-terminal extension (Dmlc2delta2-46) was approximately 1 nm less than that of wild type (48.56 +/- 0.02 nm). The myofilament lattice of detergent-treated, demembranated DLM swelled, with the DmlcdeltaA2-46 lattice expanding more than wild type and requiring roughly twice the concentration of Dextran T500 to restore its lattice to in vivo spacing (9-10% vs. 4% w/v). The calcium sensitivity and maximum amplitude of net oscillatory work near the in vivo lattice spacing was significantly lower in Dmlc2delta2-46 compared to wild type (pCa50 shifted by approximately one-third of a pCa unit; amplitude reduced by approximately one-half). These changes were in contrast to the lack of effect reported in a previous study carried out in the absence of Dextran T500. The results are consistent with the N-terminal extension interacting with actin to increase the probability that crossbridges form during stretch-activated oscillatory work and power production, especially at submaximal levels of calcium activation.
在整个动物界发现的某些II类肌球蛋白轻链N端延伸的氨基酸序列和基序的相似性表明其具有共同的功能作用。N端延伸的一个可能作用是增强通常以振荡模式运作的横纹肌中的振荡功和功率产生。我们进行了小角X射线衍射实验和小长度微扰分析,以研究删除果蝇飞行肌中肌球蛋白调节轻链(RLC)的N端延伸的结构和功能后果。缺乏RLC N端延伸(Dmlc2delta2 - 46)的果蝇背纵肌(DLM)的体内晶格间距比野生型小约1 nm(48.56 +/- 0.02 nm)。经去污剂处理、去膜的DLM的肌丝晶格膨胀,DmlcdeltaA2 - 46晶格比野生型膨胀得更多,并且需要大约两倍浓度的葡聚糖T500才能将其晶格恢复到体内间距(9 - 10% w/v对4% w/v)。与野生型相比,Dmlc2delta2 - 46在接近体内晶格间距时的钙敏感性和净振荡功的最大幅度显著降低(pCa50偏移约三分之一pCa单位;幅度降低约一半)。这些变化与之前在没有葡聚糖T500的情况下进行的一项研究中报道的无影响形成对比。结果与N端延伸与肌动蛋白相互作用以增加在拉伸激活的振荡功和功率产生过程中形成横桥的概率一致,特别是在次最大钙激活水平。