Characterization of a streptococcal endopeptidase with homology to human endothelin-converting enzyme.
Characterization of a streptococcal endopeptidase with homology to human endothelin-converting enzyme.
复制标题
与人内皮素转化酶同源的链球菌内肽酶的表征。
DOI：
10.1128/iai.69.1.58-64.2001
复制
发表时间：
2001
影响因子：
3.1
通讯作者：
Froeliger,E
中科院分区：
文献类型：
--
作者：
Oetjen,J;Fives-Taylor,P;Froeliger,E
A gene encoding an endopeptidase fromStreptococcus parasanguisFW213 has been cloned and shown to have high sequence homology to genes encoding mammalian metalloendopeptidases. The gene, designatedS. parasanguis pepO, was cloned into the pET28a expression vector, resulting in a fusion of vector sequences encoding a hexahistidine tag at the carboxyl terminus. The recombinant PepO (rPepO) was expressed inEscherichia coliand purified using an Ni2+affinity column. Polyclonal antiserum to rPepO was raised in rabbits and used to localize FW213 PepO to the cytosol. Southern hybridization and immunoblot analysis revealed that other oral streptococci contain regions of DNA with homology topepOand produce a protein with antigenic properties similar to that of FW213 PepO. Enzymatic activity assays indicated that onlyS. parasanguisspecies possess the ability to cleave metenkephalin, the natural substrate of the human neutral endopeptidase (NEP). Inhibition assays revealed thatS. parasanguisPepO is a member of the M13 category of metalloendopeptidases, which includes NEP and endothelin-converting enzyme 1 (ECE-1), an enzyme involved in the maintenance of vascular tone. Thiorphan and phosphoramidon, two specific inhibitors of this category of endopeptidases, were used to determine thatS. parasanguisPepO is more similar to ECE-1 than to NEP.
