CLONING AND NUCLEOTIDE-SEQUENCE OF OPDA, THE GENE ENCODING OLIGOPEPTIDASE-A IN SALMONELLA-TYPHIMURIUM
CLONING AND NUCLEOTIDE-SEQUENCE OF OPDA, THE GENE ENCODING OLIGOPEPTIDASE-A IN SALMONELLA-TYPHIMURIUM
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DOI：
10.1128/jb.174.5.1631-1640.1992
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发表时间：
1992-03-01
影响因子：
3.2
通讯作者：
MILLER, CG
中科院分区：
文献类型：
--
作者：
CONLIN, CA;MILLER, CG
鼠伤寒沙门氏菌（Salmonella typhimurium）的opdA基因（以前称为optA）编码金属肽酶，寡肽酶A（OpdA），首先通过其切割和允许利用N-乙酰基-L-Ala 4（E. R.维米尔湖绿色和C. G.米勒，细菌学杂志。 153：1259-1265，1983）。 分离携带opdA基因的pBR 328的衍生物，并显示其以比野生型高约100倍的水平表达寡肽酶活性。 这些质粒补充了与opdA突变相关的所有表型（未能使用N-乙酰基-L-Ala 4、缺陷的噬菌体P22发育和减弱的内肽酶活性）。 这些质粒之一（pCM 127）的opdA区由Tn 1000（γ（δ））的插入限定，并且这些插入被用作引物位点以确定插入DNA的2，843-bp片段的核苷酸序列。 该区域含有编码680个氨基酸的蛋白质的开放阅读框，其N末端与纯化的OpdA的N末端一致。 该开放阅读框包含Zn 2+金属蛋白酶的典型序列基序和推定的σ（32）启动子。 然而，通过使用β-半乳糖苷酶操纵子融合，在温度变化后未检测到诱导。 预测的OpdA序列与S.鼠伤寒沙门氏菌基因dcp和大鼠金属肽酶EC 3.4.24.15.，参与肽激素的加工。
The opdA gene (formerly called optA) of Salmonella typhimurium encodes a metallopeptidase, oligopeptidase A (OpdA), first recognized by its ability to cleave and allow utilization of N-acetyl-L-Ala4 (E. R. Vimr, L. Green, and C. G. Miller, J. Bacteriol. 153:1259-1265, 1983). Derivatives of pBR328 carrying the opdA gene were isolated and shown to express oligopeptidase activity at levels approximately 100-fold higher than that of the wild type. These plasmids complemented all of the phenotypes associated with opdA mutations (failure to use N-acetyl-L-Ala4, defective phage P22 development, and diminished endopeptidase activity). The opdA region of one of these plasmids (pCM127) was defined by insertions of Tn1000 (gamma(delta)), and these insertions were used as priming sites to determine the nucleotide sequence of a 2,843-bp segment of the insert DNA. This region contained an open reading frame coding for a 680-amino-acid protein, the N terminus of which agreed with that determined for purified OpdA. This open reading frame contained both a sequence motif typical of Zn2+ metalloproteases and a putative sigma(32) promoter. However, no induction was detected upon temperature shift by using a beta-galactosidase operon fusion. The predicted OpdA sequence showed similarity to dipeptidyl carboxypeptidase, the product of the S. typhimurium gene dcp, and to rat metallopeptidase EC 3.4.24.15., which is involved in peptide hormone processing.