Saito T, Ishihara K, Kato T, Okuda K
Research & Development Department, Oral Care Business Headquarters, Sunstar Inc., Takatsuki, Osaka, Japan.
Infect Immun. 1997 Nov;65(11):4888-91. doi: 10.1128/iai.65.11.4888-4891.1997.
We have isolated and characterized an N-benzoyl-Val-Gly-Arg-p-nitroanilide-specific protease gene, designated prtH, from Bacteroides forsythus ATCC 43037. Nucleotide sequencing of the DNA insert from the clone (hereafter referred to as clone FST) revealed that the protease activity corresponded to an open reading frame consisting of 1,272 bp coding for a 47.8-kDa protein. When plasmid pFST was used as a probe in Southern hybridization, Sau3AI-digested chromosomal DNA of B. forsythus ATCC 43037 as well as the chromosomal DNAs of the isolated strains Ta4, TR5, and YG2 showed 0.6- and 0.8-kb hybridizing bands. The cell-free extracts of clone FST showed hemolytic activity on human blood cells. The hydrolytic activity of cell extracts of the pFST clone was inhibited by p-toluenesulfonyl-L-lysine chloromethyl ketone hydrochloride, leupeptin, N-ethylmaleimide, iodoacetic acid, iodoaceteamide, and EDTA.
我们从福赛斯坦纳菌ATCC 43037中分离并鉴定了一个N-苯甲酰基-Val-Gly-Arg-对硝基苯胺特异性蛋白酶基因，命名为prtH。对克隆（以下简称克隆FST）中DNA插入片段的核苷酸测序显示，蛋白酶活性对应于一个由1272 bp组成的开放阅读框，编码一个47.8 kDa的蛋白质。当质粒pFST用作Southern杂交的探针时，福赛斯坦纳菌ATCC 43037的Sau3AI消化的染色体DNA以及分离菌株Ta4、TR5和YG2的染色体DNA显示出0.6 kb和0.8 kb的杂交带。克隆FST的无细胞提取物对人血细胞具有溶血活性。pFST克隆的细胞提取物的水解活性受到对甲苯磺酰-L-赖氨酸氯甲基酮盐酸盐、亮抑蛋白酶肽、N-乙基马来酰亚胺、碘乙酸、碘乙酰胺和EDTA的抑制。
