竹叶青蛇毒凝血酶样酶氨基酸序列报道  

Primary Amino Acid Sequence of a Thrombin like Enzyme from the Venom of Trimeresurus Stejnegeri

作　　者：张云[1] 李文辉[1] 高荣[1] 熊郁良[1] 王婉瑜[1] 

机构地区：[1]中国科学院昆明动物研究所

出　　处：《中国生物化学与分子生物学报》1998年第1期77-81,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：中国科学院院长基金;云南省科委主任基金

摘　　要：蛇毒凝血酶样酶可作为蛋白酶结构与功能研究的良好模型，并已广泛用于各种血栓疾病的诊断和治疗，因而测定其一级结构具有重要意义．利用逆转录与聚合酶链反应相结合的ＲＴ－ＰＣＲ法，扩增出竹叶青蛇毒凝血酶样酶（ＴＳＶ－ＴＥＬ１）的ｃＤＮＡ；将扩增的ｃＤＮＡ片段克隆入ｐＧＥＭ－Ｔ载体中，经末端终止法测定核苷酸序列，推导出竹叶青蛇毒凝血酶样酶的全序列．竹叶青蛇毒凝血酶样酶由２３４个氨基酸组成并含有１个位于Ａｓｎ２０的Ｎ－型糖基结合位点．竹叶青蛇毒凝血酶样酶序列与其它蛇种来源凝血酶样酶具有较大相似性，其与黄绿烙铁头蛇毒凝血酶样酶序列相似度为８４％，与美洲矛头蝮蛇毒凝血酶样酶序列相似度为６８％，而与牛凝血酶Ｂ链序列相似度仅为２５％．Snake venom thrombin like enzymes are good models for the study of structure function relationship of proteases.They are also widely used in the diagnosis and treatment of thrombosis in clinic.It is important to determine their amino acid sequences.The cDNA encoding Trimeresurus stejnegeri venom thrombin like enzyme (TSV TLE1) was amplified by reverse transcription polymerase chain reaction(RT PCR)method.The amplified cDNA was subcloned by the deoxythymidine tailed vector method into a pGEM T plasmid.The nucleotide sequence of the cDNA was determined.The deduced complete amino acid sequence of TSV TLE1 indicates that it is composed of 234 amino acids and contains a potential N glycosylation site at Asn 20 .The sequence of TSV TLE1 exhibits a high degree of sequence identity with other venom thrombin like enzymes,84% with flavoxobin from Trimeresurus flavoviridis venom,68% with batroxobin from Bothrops atrox venom.On the other hand,TSV TLE1 only shows 25% sequence identity with bovine thrombin and 42% with rat trypsin.

关 键 词：竹叶青 蛇毒 蛋白酶 凝血酶 氨基酸序列 

分 类 号：Q556[生物学—生物化学]