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Cloning and sequencing of cDNA for fish slow skeletal muscle myosin light chains

鱼类慢骨骼肌肌球蛋白轻链 cDNA 的克隆和测序

基本信息

批准号：
15580180
负责人：
ISHIZAKI Shoichiro
金额：
$ 1.54万
依托单位：
Tokyo University of Marine Science and Technology (2004)東京水産大学 (2003)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15580180/
关键词：
slow skeletal muscle myosin light chain cDNA cloning fish 熱安定性

项目摘要

Various species of fish have two types of skeletal muscle, the ordinary (white) and dark (red) muscles. It has been suggested that ordinary muscle corresponds to fast muscle of higher vertebrates, while dark muscle corresponds to slow muscle. Both skeletal muscle myosins consist of two heavy chains of approximately 200kDa and four light chains of approximately 20kDa as in the case of higher-vertebrate counterparts. The fish fast skeletal muscle myosin has been shown to have three kinds of light chains, which are classified into two groups, one pair of alkali light chains consisting of two heterologous components (A1 and A2 in the order of decreasing molecular weight) and another pair of 5,5'-dithio-bis-2-nitrobenzoic acid (DTNB) light chains. On the other hand, it is well-known that fish slow muscle myosin differs from fast muscle myosin in light chain composition. The former myosin generally has only two kinds of light chain (D1 and D2) as does cardiac myosin. However, information on structural characterization of fish slow skeletal muscle myosin light chains is not available at all. This study described cloning, sequencing, and characterization of cDNA clones encoding D1 and D2 light chains of striped jack Caranx delicatissimus and horse mackerel Trachurus japonicus slow skeletal muscles, respectively. D1 light chain was found to have a part of the difference peptide portion in the N-terminal region, as was detected in A1 light chains. This result strongly suggests that D1 light chain in slow skeletal muscle myosin possess the same function as A1 light chain in fast skeletal muscle myosin. D2 light chain was also found to have serine residue, showing phosphorylation as was detected in DTNA light chain of fast muscle. The results in this study were the first known reports on the complete primary structure of fish slow muscle myosin light chains.

不同种类的鱼有两种类型的骨骼肌，普通（白色）和深色（红色）肌肉。有人认为，普通肌肉对应于高等脊椎动物的快肌，而暗肌对应于慢肌。两种骨骼肌肌球蛋白都由两条约200 kDa的重链和四条约20 kDa的轻链组成，与高等脊椎动物对应物的情况相同。鱼类快速骨骼肌肌球蛋白具有三种轻链，分为两组，一对由两种异源组分（A1和A2，按分子量递减的顺序）组成的碱性轻链和另一对5，5 '-二硫代-双-2-硝基苯甲酸（DTNB）轻链。另一方面，众所周知，鱼慢肌肌球蛋白在轻链组成上不同于快肌肌球蛋白。前者肌球蛋白通常只有两种轻链（D1和D2），心脏肌球蛋白也是如此。然而，鱼慢骨骼肌肌球蛋白轻链的结构特征的信息是不可用的。本研究描述了克隆，测序和表征的cDNA克隆编码D1和D2轻链的条纹杰克鱼delicatissimus和竹荚鱼Trachurus的骨骼肌，分别。发现D1轻链在N-末端区域具有部分差异肽部分，如在A1轻链中检测到的。这一结果有力地表明，骨骼肌慢肌球蛋白中的D1轻链具有与骨骼肌快肌球蛋白中的A1轻链相同的功能。还发现D2轻链具有丝氨酸残基，显示磷酸化，如在快肌的DTNA轻链中检测到的。这项研究的结果是第一次报道鱼类慢肌肌球蛋白轻链的完整一级结构。