摘要
组织蛋白酶L在多种生理过程中具有重要作用。根据本实验室构建的三角帆蚌(Hyriopsis cummgii)cDNA文库中已标注的EST序列,利用RACE方法克隆了三角帆蚌组织蛋白酶L基因的cDNA全序列(GenBank登录号为HQ610996)。结果表明,该序列全长为1 105 bp,包括24 bp的5′-末端非转录区,1 002 bp的开放阅读框和79 bp的3′-末端非转录区,共编码333个氨基酸,包含1个由20个氨基酸组成的信号肽。推测的三角帆蚌组织蛋白酶L前体肽具有组织蛋白酶前体抑制因子功能结构域,具有组织蛋白酶L家族高度保守结构基序ERFNIN[E-X3-R-X2-(L/V)-F-X3-N-X3-I-X3-N]、GNFD和GCXGG;该蛋白的半胱氨酸类蛋白酶半胱氨酸、组氨酸和天冬氨酸活性位点均高度保守。同源性分析表明,推测的三角帆蚌组织蛋白酶L氨基酸序列与其他贝类高度保守,同源性在60%-74%之间。进化分析显示,三角帆蚌组织蛋白酶L与其他贝类组织蛋白酶L聚为一支,在该支中,三角帆蚌与软体动物淡水螺(R.peregra)的亲缘关系最近。本研究结果为进一步研究三角帆蚌组织蛋白酶L的生理功能提供基础资料。
Cathepsin L plays important roles in many physiological processes. In this study, according to the EST sequence from cDNA library of Hyriopsis cumingii,a cDNA encoding cathepsin L（ GenBank accession number： HQ610996）was identified and charac- terized from Hyriopsis cumingii by RACE method. The cathepsin L gene cDNA consisted of 1 105 bp with an open reading frame（ORF） of 1 002 bp, a 24 bp 5＇-untranslated region, and a 79 bp 3 ＇-untranslated region. The Open reading frame encoded a protein of 333 amino acids with 20 residues signal sequence. The H. cumingii cathepsin L amino acids sequence was predicted to contain a cathepsin propep- tide inhibitor domain, and three motifs, ERFNIN, GNFD and GCXGG. ERFNIN motif is a highly conserved motif that is interspersed throughout the cathepsin L family. The cysteine active site,histidine active site and asparagine active site of cathepsin L of H. cumingii were also highly conserved. Homology analysis of cathepsin L revealed that the cathepsin L of H. cumingii shared 60% - 74% identity to other known cathepsin L sequences. The phylogenetic tree showed that cathepsin L of 11. cumingii clustered with cathepsin L of other molluscs and was closely related to cathepsin L of Radix peregra. These results provided preliminary information for further exploring the roles of cathepsin L of H. cumingii.
出处
《生物技术通报》
CAS
CSCD
北大核心
2011年第6期104-111,共8页
Biotechnology Bulletin
基金
"973"计划前期研究专项项目(2009CB126001)
上海市科委地方院校能力建设项目(08390510100)
国家自然科学基金青年科学基金项目(31001110)
上海市水产养殖重点学科建设项目(Y1101)
