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

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetyltransferases*
  • Amino Acid Sequence
  • Antibodies, Monoclonal / immunology
  • Bacterial Proteins / biosynthesis*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / immunology
  • Bacterial Proteins / metabolism
  • Base Sequence
  • Bleomycin / biosynthesis
  • Bleomycin / metabolism*
  • Bleomycin / pharmacology
  • Blotting, Western
  • Carrier Proteins / biosynthesis*
  • Carrier Proteins / genetics
  • Carrier Proteins / immunology
  • Carrier Proteins / metabolism
  • Escherichia coli / metabolism
  • Isoelectric Point
  • Methicillin Resistance
  • Molecular Sequence Data
  • Promoter Regions, Genetic
  • Recombinant Proteins / biosynthesis
  • Staphylococcus aureus / drug effects
  • Staphylococcus aureus / metabolism*
  • Streptomyces / metabolism*

Substances

  • Antibodies, Monoclonal
  • Bacterial Proteins
  • Carrier Proteins
  • Recombinant Proteins
  • Bleomycin
  • bleomycin N-acetyltransferase
  • Acetyltransferases