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

[摘要]

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-chracterize 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.

[学科分类] 生物化学/生物物理

[关键词] Bleomycin;Bleomycin-binding protein;Methicillin-resistant Staphylococcus aureus;Monoclonal antibody;Polymerase chain reaction;Streptomyces verticillus;Ap;ampicillin;Bm;bleomycin;blmA;gene encoding Bm-binding protein from Streptomyces verticillus;BLMA;protein encoded by blmA;blmS;gene encoding bleomycin-binding protein from a methicillin resistant Staphylococcus aureus;BLMS;a protein by blmS;IPTG;isopropyl-β-d-thiogalactoside;malE;gene encoding MBP;MBP;maltose-binding protein;MBP/BLMA;a fusion protein between MBP and BLMA;MRSA;methicillin resistant Staphylococcus aureus;PAGE;polyacrylamide gel electrophoresis;SDS;sodium dodecyl sulfate