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浙江大学学报（农业与生命科学版）
浙江大学学报(农业与生命科学版)  2012, Vol. 38 Issue (4): 370-376    DOI: 10.3785/j.issn.1008-9209.2012.04.003
生物科学与技术     
总状毛霉丝氨酸蛋白酶基因结构和功能的生物信息学分析
柯野， 曾松荣， 郑秋桦
韶关学院 英东生命科学学院，广东 韶关 512005
Bioinformatics analysis of structure and function of serine protease gene from Mucor racemosus
KE Ye, ZENG Song-rong, ZHENG Qiu-hua
Yingdong College of Life Sciences, Shaoguan University, Shaoguan, Guangdong 512005, China
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摘要： 利用 PCR和RACE(cDNA末端快速扩增)技术从总状毛霉中克隆获得1个丝氨酸蛋白酶新基因，利用生物信息学方法对该基因序列、编码的酶序列以及酶性质和结构进行预测。结果表明：该基因编码区序列长1421 bp，由3个外显子和2个内含子组成，编码由429个氨基酸残基组成的多肽；对该多肽的组成和进化分析表明，其成熟肽属于蛋白酶K家族的胞外蛋白酶；通过同源建模结构分析表明，该酶没有二硫键，有152个氢键和29个盐键，酶的Glu190、Ala192和Asp215残基与一个Ca2+相结合，酶活性位点残基Asp44和Ser241溶剂可及表面积较大，分别达到0.2114 2和3.3093 2；该酶的底物结合区域中，S1和S4口袋结构明显，S2次之，S3最不明显，部分组成S1和S4口袋的氨基酸残基不保守，这可能使该酶具有独特的水解性。上述结果可为该蛋白酶基因的异源表达和结构与功能关系等方面的研究提供参考。
Abstract: By PCR and RACE (rapid amplification of cDNA ends) technique, a new serine protease gene was obtained from Mucor racemosus. The gene sequence, the putative protease sequence, characteristics and structure were predicted and analyzed by bioinformatics method. The results proved that the length of gene coding region was 1421 bp, which contained 3 exons and 2 introns, and this gene encoded a peptide chain with 429 amino acids. The property and evolutionary relationship of the peptide chain were predicted. As a result, the mature peptide was a kind of secreted protease, which belonged to proteinase K family. Structural models of the protease were modeled by the homology modeling technique. The results showed that it had no disulfide bridge, contained 152 hydrogen bonds, 29 salt bonds, and the residues Glu190, Ala192 and Asp215 binding with a calcium ion. Solvent accessible surface of Asp44 and Ser241, which were active site residues, were very high, and respectively reached to 0.2114 2 and 3.3093 2. For substrate binding regions, S1 and S4 were distinct and large pockets, and S2 was  less distinct and smaller, and S3 was not  distinct. Some residues forming S1 and S4 were not very conservative, so the change of residues propably resulted in special hydrolysis of the protease. Results above provide  a further basis for studying the expression of transgenes and relationship between structure and function for  the serine protease.
出版日期: 2012-07-25
CLC:  Q 816  
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柯野
曾松荣
郑秋桦

引用本文:

柯野， 曾松荣， 郑秋桦. 总状毛霉丝氨酸蛋白酶基因结构和功能的生物信息学分析[J]. 浙江大学学报(农业与生命科学版), 2012, 38(4): 370-376.

KE Ye, ZENG Song-rong, ZHENG Qiu-hua. Bioinformatics analysis of structure and function of serine protease gene from Mucor racemosus. , 2012, 38(4): 370-376.

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http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2012.04.003    或    http://www.zjujournals.com/agr/CN/Y2012/V38/I4/370

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