CAJ | 학술논문

从菜花烙铁头蛇的毒腺中,利用RT-PCR进行体外扩增,克隆到2个金属蛋白酶-去整合素基因,命名为TJM-l、TJM-2.TJM-1 cDNA全长为1 528 bp,编码481个氨基酸;TJM-2 cDNA全长为1 578bp,编码484个氨基酸.TJM-1和TJM-2都属于Ⅱ型蛇毒金属蛋白酶,由信号肽、前肽、金属蛋白酶、间隔肽和去整合素5部分组成.Ⅱ型蛇毒金属蛋白酶氨基酸序列的比较及进化分析显示,它可进一步分为两类,一类包括大多数Ⅱ型蛇毒金属蛋白酶(其中含有TJM-1),而TJM-2和agkistin则组成了另一类.并且TJM-2和agkistin的第407位和第426位残基都是半胱氨酸,而在其它Ⅱ型金属蛋白酶的相应位置,407位是丝氨酸,426位则缺失.TJM-2和agkistin均有可与整合素α2I区域特异性结合的片段Q/NRKRHDNAQ(残基276～284),这个片段在其它Ⅱ型金属蛋白酶中并没有发现.因此推断,TJM-2和agkistin可能属于一类新型的Ⅱ型蛇毒金属蛋白酶.
종채화락철두사적독선중,이용RT-PCR진행체외확증,극륭도2개금속단백매-거정합소기인,명명위TJM-l、TJM-2.TJM-1 cDNA전장위1 528 bp,편마481개안기산;TJM-2 cDNA전장위1 578bp,편마484개안기산.TJM-1화TJM-2도속우Ⅱ형사독금속단백매,유신호태、전태、금속단백매、간격태화거정합소5부분조성.Ⅱ형사독금속단백매안기산서렬적비교급진화분석현시,타가진일보분위량류,일류포괄대다수Ⅱ형사독금속단백매(기중함유TJM-1),이TJM-2화agkistin칙조성료령일류.병차TJM-2화agkistin적제407위화제426위잔기도시반광안산,이재기타Ⅱ형금속단백매적상응위치,407위시사안산,426위칙결실.TJM-2화agkistin균유가여정합소α2I구역특이성결합적편단Q/NRKRHDNAQ(잔기276～284),저개편단재기타Ⅱ형금속단백매중병몰유발현.인차추단,TJM-2화agkistin가능속우일류신형적Ⅱ형사독금속단백매.
Two cDNAs encoding metalloproteinase-disintegrin, named TJM-1 and TJM-2, were cloned from venom gland of Trimeresurus jerdonii by reverse transcription polymerase chain reaction (RT-PCR). The fulllength cDNA of TJM-1 was 1 528 bp and encoded an open reading frame (ORF) of 481 amino acids. That of TJM-2 was 1578 bp and encoded an ORF of 484 amino acids. Both encoding proteins belonged to the class Ⅱ of snake venom metallproteinases (SVMP) (class P-Ⅱ), consisting of signal peptide sequence, zymogen sequence, metalloproteinase, spacer and disintegrin domains. Phylogenetic analysis of class P-Ⅱ SVMPs revealed it could be further classified into two groups. One group contained most class P-Ⅱ SVMPs including TJM-I. TJM-2 and Agkistin formed the other group. Moreover, the 407th and 426th residues in TJM-2 and agkistin were Cys, while in other class P-Ⅱ metalloproteinases 407th residue was Ser and 426th residue was deleted. The segments of Q/NRKRHDNAQ (residue 280-288) of TJM-2 and agkistin, which were thought to specifically bind to integrin α2 I domain, was also not found in class P- Ⅱ SVMPs. It was predicted that TJM2 and agkistin might belong to a new type of class P- Ⅱ SVMPs.