Tanokura M, Tawada Y, Ohtsuki I
J Biochem. 1982 Apr;91(4):1257-65. doi: 10.1093/oxfordjournals.jbchem.a133810.
Chymotryptic subfragments from rabbit skeletal troponin T were purified using column chromatography. Molecular weight values on SDS gel electrophoresis, tryptophan contents, N- and C-terminal residues, and amino acid compositions were examined for each subfragment. Based on these findings, the positions of the subfragments in the sequence of troponin T were determined as follows: N-terminal acetylserine-1-tyrosine-158 for troponin T1 (MW 18,700); serine-156-C-terminal lysine-259 for troponin T2 alpha s (MW 12,200); leucine-159-C-terminal lysine-259 for troponin T2 (or troponin T2 alpha) (MW 11,900); leucine-159-phenylalanine-242 for troponin T2 beta I (MW 10,200); leucine-159-tyrosine-227 for troponin T2 beta II (MW 8,400); leucine-159-leucine-222 for troponin T2 beta III (MW 7,700); and serine-243-C-terminal lysine-259 for troponin T2 gamma (MW 1,800). The pathway of chymotryptic digestion of troponin T was also investigated and the results are discussed in relation to the higher structure of troponin T.l The interaction of some chymotryptic subfragments with tropomyosin was also investigated by affinity chromatography.
使用柱色谱法纯化了来自兔骨骼肌肌钙蛋白T的胰凝乳蛋白酶亚片段。对每个亚片段进行了SDS凝胶电泳的分子量值、色氨酸含量、N端和C端残基以及氨基酸组成的检测。基于这些发现，肌钙蛋白T序列中亚片段的位置确定如下：肌钙蛋白T1（分子量18,700）的N端乙酰丝氨酸-1-酪氨酸-158；肌钙蛋白T2αs（分子量12,200）的丝氨酸-156-C端赖氨酸-259；肌钙蛋白T2（或肌钙蛋白T2α）（分子量11,900）的亮氨酸-159-C端赖氨酸-259；肌钙蛋白T2βI（分子量10,200）的亮氨酸-159-苯丙氨酸-242；肌钙蛋白T2βII（分子量8,400）的亮氨酸-159-酪氨酸-227；肌钙蛋白T2βIII（分子量7,700）的亮氨酸-159-亮氨酸-222；以及肌钙蛋白T2γ（分子量1,800）的丝氨酸-243-C端赖氨酸-259。还研究了肌钙蛋白T的胰凝乳蛋白酶消化途径，并结合肌钙蛋白T的高级结构对结果进行了讨论。还通过亲和色谱法研究了一些胰凝乳蛋白酶亚片段与原肌球蛋白的相互作用。
