来自博来霉素产生菌轮枝链霉菌和耐甲氧西林金黄色葡萄球菌的博来霉素结合蛋白在大肠杆菌中的过量表达及其免疫学特性分析

Overproduction of the bleomycin-binding proteins from bleomycin-producing Streptomyces verticillus and a methicillin-resistant Staphylococcus aureus in Escherichia coli and their immunological characterisation.

作者信息

Sugiyama M, Kumagai T, Matsuo H, Bhuiyan M Z, Ueda K, Mochizuki H, Nakamura N, Davies J E

机构信息

Institute of Pharmaceutical Sciences, Hiroshima University School of Medicine, Japan.

出版信息

FEBS Lett. 1995 Mar 27;362(1):80-4. doi: 10.1016/0014-5793(95)00218-x.

DOI:10.1016/0014-5793(95)00218-x

PMID:7535252

Abstract

The bleomycin-binding proteins designated BLMA and BLMS, which confer resistance to bleomycin (Bm), from Bm-producing Streptomyces verticillus ATCC15003 and a methicillin-resistant Staphylococcus aureus B-26, respectively, were overexpressed in Escherichia coli. The present study showed that both BLMA and BLMS quench the antibacterial activity of Bm by the binding to the drug. To immuno-characterize the Bm-binding proteins, we constructed a monoclonal antibody against BLMA. The antibody, designated 893-12, did not cross react to BLMS and another Bm-binding protein from tallysomycin-producing Streptoalloteichus hindustanus. Although the ability of Bm to cleavage DNA was eliminated by a binding of BLMA to Bm, as shown by Sugiyama et al. [Gene 151 (1994) 11-15], the Bm-induced DNA degradation was restored by pre-incubation of BLMA with the anti-BLMA monoclonal antibody.

摘要

分别来自产博来霉素的轮状链霉菌ATCC15003和耐甲氧西林金黄色葡萄球菌B-26的赋予对博来霉素（Bm）抗性的博来霉素结合蛋白BLMA和BLMS在大肠杆菌中过表达。本研究表明，BLMA和BLMS均通过与药物结合来淬灭Bm的抗菌活性。为了对Bm结合蛋白进行免疫特性分析，我们构建了一种针对BLMA的单克隆抗体。该抗体命名为893-12，与BLMS以及来自产 tallysomycin的印度链霉菌的另一种Bm结合蛋白无交叉反应。尽管如杉山等人[《基因》151(1994)11 - 15]所示，BLMA与Bm结合消除了Bm切割DNA的能力，但通过将BLMA与抗BLMA单克隆抗体预孵育，Bm诱导的DNA降解得以恢复。