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非洲鲶鱼(Clarias gariepinus)中肌肉生长抑制素(MSTN)的cDNA序列及蛋白质生物信息学分析

cDNA sequence and protein bioinformatics analyses of MSTN in African catfish (Clarias gariepinus).

作者信息

Kanjanaworakul Poonmanee, Sawatdichaikul Orathai, Poompuang Supawadee

机构信息

Center for Agricultural Biotechnology, Kasetsart University, Nakorn Pathom, 73140, Thailand.

Center of Excellence on Agricultural Biotechnology (AG-BIO/PERDO-CHE), Bangkok, 10900, Thailand.

出版信息

Mol Biol Rep. 2016 Apr;43(4):283-93. doi: 10.1007/s11033-016-3961-7. Epub 2016 Feb 24.

Abstract

Myostatin, also known as growth differentiation factor 8, has been identified as a potent negative regulator of skeletal muscle growth. The purpose of this study was to characterize and predict function of the myostatin gene of the African catfish (Cg-MSTN). Expression of Cg-MSTN was determined at three growth stages to establish the relationship between the levels of MSTN transcript and skeletal muscle growth. The partial cDNA sequence of Cg-MSTN was cloned by using published information from its congener walking catfish (Cm-MSTN). The Cg-MSTN was 1194 bp in length encoding a protein of 397 amino acids. The deduced MSTN sequence exhibited key functional sites similar to those of other members of the TGF-β superfamily, especially, the proteolytic processing site (RXXR motif) and nine conserved cysteines at the C-terminal. Expression of MSTN appeared to be correlated with muscle development and growth of African catfish. Protein bioinformatics revealed that the primary sequence of Cg-MSTN shared 98 % sequence identity with that of walking catfish Cm-MSTN with only two different residues, [Formula: see text]. and [Formula: see text]. The proposed model of Cg-MSTN revealed the key point mutation [Formula: see text] causing a 7.35 Å shorter distance between the N- and C-lobes and an approximately 11° narrow angle than those of Cm-MSTN. The substitution of a proline residue near the proteolytic processing site which altered the structure of myostatin may play a critical role in reducing proteolytic activity of this protein in African catfish.

摘要

肌肉生长抑制素,也被称为生长分化因子8,已被确定为骨骼肌生长的一种强效负调节因子。本研究的目的是对非洲鲶鱼(Cg-MSTN)的肌肉生长抑制素基因的功能进行表征和预测。在三个生长阶段测定Cg-MSTN的表达,以建立肌肉生长抑制素转录水平与骨骼肌生长之间的关系。利用其同属物种步行鲶鱼(Cm-MSTN)已发表的信息克隆了Cg-MSTN的部分cDNA序列。Cg-MSTN长度为1194 bp编码一个397个氨基酸的蛋白质。推导的肌肉生长抑制素序列显示出与转化生长因子-β超家族其他成员相似的关键功能位点,特别是蛋白水解加工位点(RXXR基序)和C端的9个保守半胱氨酸。肌肉生长抑制素的表达似乎与非洲鲶鱼的肌肉发育和生长相关。蛋白质生物信息学显示,Cg-MSTN的一级序列与步行鲶鱼Cm-MSTN的序列有98%的同一性,只有两个不同的残基,[公式:见正文]和[公式:见正文]。所提出的Cg-MSTN模型揭示了关键的点突变[公式:见正文],导致N叶和C叶之间的距离比Cm-MSTN短7.35 Å,角度窄约11°。蛋白水解加工位点附近脯氨酸残基的取代改变了肌肉生长抑制素的结构,可能在降低非洲鲶鱼中该蛋白的蛋白水解活性方面起关键作用。

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