Purification and Characterization of Virginiae Butanolide C-binding Protein, a Possible Pleiotropic Signal-Transducer in Streptomyces Virginiae

J Antibiot (Tokyo). 1990 Jun;43(6):692-706. doi: 10.7164/antibiotics.43.692.

Abstract

Virginiae butanolide C (VB-C) is an autoregulator which triggers virginiamycin production in Streptomyces virginiae. A new binding assay with tritium-labeled VB-C analogue (2,3-cis-2-(1'-hydroxy-[6',7'-3H]heptyl)-3-(hydroxymethyl)butanolide+ ++ ) was developed and a specific VB-C binding protein was purified to homogeneity from crude extracts of S. virginiae by ammonium sulfate fractionation, DEAE-Sephacel and Sephadex G-100 column chromatographies, hydrophobic HPLC on phenyl 5PW and native polyacrylamide gel electrophoresis. The VB-C binding protein showed an apparent Mr of 35,800 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Mr of 26,000 approximately 44,000 on native molecular sieve HPLC, indicating the monomeric nature of the binding protein. The binding protein efficiently bound to a VB affinity column and eluted specifically by VB-C, which confirmed the specific nature of the binding protein. The binding activity decreased by 40% in the presence of genomic DNA from S. virginiae, indicating interaction between the VB-C binding protein and the DNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 4-Butyrolactone / analogs & derivatives
  • 4-Butyrolactone / metabolism*
  • Carrier Proteins / analysis
  • Carrier Proteins / isolation & purification*
  • Chemical Phenomena
  • Chemical Precipitation
  • Chemistry
  • Chromatography, Affinity
  • Chromatography, High Pressure Liquid
  • DNA, Bacterial / pharmacology
  • Densitometry
  • Electrophoresis, Polyacrylamide Gel
  • Furans / metabolism*
  • Ligands
  • Molecular Structure
  • Signal Transduction*
  • Streptomyces / genetics
  • Streptomyces / metabolism*

Substances

  • Carrier Proteins
  • DNA, Bacterial
  • Furans
  • Ligands
  • virginiamycin butanolide C
  • 4-Butyrolactone