Rindt H, Bauer B J, Robbins J
Department of Pharmacology and Cell Biophysics, University of Cincinnati, College of Medicine, OH 45267.
J Muscle Res Cell Motil. 1993 Feb;14(1):26-34. doi: 10.1007/BF00132177.
In order to initiate studies on the structural and functional relationships of the myosin heavy chain, we constructed a full-length complementary DNA encoding the isoform that is found in the fast white muscle of the embryonic chicken. The complementary DNA contained 108 basepairs of its 3'-untranslated region and was preceded by a leader sequence derived from the alfalfa mosaic virus. Similarly, a complementary DNA encoding 963 amino acids which encompass the subfragment-1 of myosin and part of the subfragment-2 was also constructed. Each was inserted into the expression vector pMT2 and transiently transfected into COS-1 cells. Both constructs directed the expression of the respective proteins, each of which was immunogenic. The full-length and subfragment-1 proteins interacted with actin and demonstrated high levels of a K(+)-activated, EDTA-resistant ATPase activity, which is characteristic of myosin.
为了开展关于肌球蛋白重链结构与功能关系的研究，我们构建了一个全长互补DNA，其编码在胚胎鸡的快白肌中发现的同种型。该互补DNA在其3'非翻译区含有108个碱基对，并且前面有一个源自苜蓿花叶病毒的前导序列。同样，还构建了一个编码963个氨基酸的互补DNA，这些氨基酸包含肌球蛋白的亚片段-1和部分亚片段-2。每个都插入到表达载体pMT2中，并瞬时转染到COS-1细胞中。两种构建体都指导了各自蛋白质的表达，每种蛋白质都具有免疫原性。全长和亚片段-1蛋白质与肌动蛋白相互作用，并表现出高水平的K(+)激活、EDTA抗性ATP酶活性，这是肌球蛋白的特征。
