Maita T, Yajima E, Nagata S, Miyanishi T, Nakayama S, Matsuda G
Department of Biochemistry, Nagasaki University School of Medicine.
J Biochem. 1991 Jul;110(1):75-87. doi: 10.1093/oxfordjournals.jbchem.a123546.
In the preceding paper [Maita, T., Miyanishi, T., Matsuzono, K., Tanioka, Y., & Matsuda, G. (1991) J. Biochem. 110, 68-74], we reported the amino-terminal 837-residue sequence of the heavy chain of adult chicken pectoralis muscle myosin. This paper describes the carboxyl terminal 1,097-residue sequence and the linkage of the two sequences. Rod obtained by digesting myosin filaments with alpha-chymotrypsin was redigested with the protease at high KCl concentration, and two fragments, subfragment-2 and light meromyosin, were isolated and sequenced by conventional methods. The linkage of the two fragments was deduced from the sequence of an overlapping peptide obtained by cleaving the rod with cyanogen bromide. The rod contained 1,039 amino acid residues, but lacked the carboxyl-terminal 58 residues of the heavy chain. A carboxyl-terminal 63-residue peptide obtained by cleaving the whole heavy chain with cyanogen bromide was sequenced. Thus, the carboxyl terminal 1,097-residue sequence of the heavy chain was completed. The linkage of subfragment-1 and the rod was deduced from the sequence of an overlapping peptide between the two which was obtained by cleaving heavy meromyosin with cyanogen bromide. Comparing the sequence of the adult myosin thus determined with that of chicken embryonic myosin reported by Molina et al. [Molina, M.I., Kropp, K.E., Gulick, J., & Robbins, J. (1987) J. Biol. Chem. 262, 6478-6488], we found that the sequence homology is 94%.
在前一篇论文中[Maita, T., Miyanishi, T., Matsuzono, K., Tanioka, Y., & Matsuda, G. (1991) J. Biochem. 110, 68 - 74]，我们报道了成年鸡胸肌肌球蛋白重链的氨基末端837个残基的序列。本文描述了羧基末端1097个残基的序列以及这两个序列的连接情况。用α-胰凝乳蛋白酶消化肌球蛋白丝得到的杆状部分在高KCl浓度下用该蛋白酶再次消化，分离出两个片段，即亚片段-2和轻酶解肌球蛋白，并通过常规方法进行测序。通过用溴化氰切割杆状部分得到的重叠肽段的序列推导出这两个片段的连接情况。杆状部分含有1039个氨基酸残基，但缺少重链羧基末端的58个残基。对用溴化氰切割整个重链得到的羧基末端63个残基的肽段进行了测序。因此，完成了重链羧基末端1097个残基的序列测定。通过用溴化氰切割重酶解肌球蛋白得到的亚片段-1和杆状部分之间的重叠肽段的序列推导出它们的连接情况。将如此确定的成年肌球蛋白序列与Molina等人报道的鸡胚胎肌球蛋白序列[Molina, M.I., Kropp, K.E., Gulick, J., & Robbins, J. (1987) J. Biol. Chem. 262, 6478 - 6488]进行比较，我们发现序列同源性为94%。
