Isolation and characterisation of a vitronectin-binding surface protein from Staphylococcus aureus

Biochim Biophys Acta. 1995 Jul 3;1250(1):110-6. doi: 10.1016/0167-4838(95)00076-7.

Abstract

In a previous study we demonstrated that cells of Staphylococcus aureus strain V8 bind 125I-labelled vitronectin in a receptor-ligand type of interaction, and a protein having a molecular mass of 60 kDa was identified as a putative high-affinity staphylococcal vitronectin-binding protein (Liang, O.D. et al. (1993) Biochim. Biophys. Acta 1225, 57-63). In the present communication we report on the isolation and preliminary characterisation of the 60 kDa vitronectin-binding protein. The bacterial cell surface proteins were released by stirring bacteria with 1 M LiCl at 37 degrees C for 2 h and separated on an FPLC Mono-Q column with a gradient of 0-0.5 M NaCl in 20 mM Tris buffer at pH 9.0. Fractions containing vitronectin-binding activity, assayed on microtiter plates with immobilised human vitronectin, were collected and SDS-PAGE analysis showed the content to be a single protein band at the 60 kDa position. In Western blot experiments the protein transblotted onto nitrocellulose membranes could bind soluble vitronectin. Its amino-terminal amino acid sequences showed a striking similarity with those of a 60 kDa heparan sulfate-binding protein from the same staphylococcal strain (Liang, O.D. et al. (1992) Infect. Immun. 60, 899-906), suggesting that they are identical molecules. This was supported by ligand blotting experiments where both vitronectin and heparan sulfate were shown to bind to the same protein band in parallel strips.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / analysis
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / isolation & purification*
  • Blotting, Western
  • Chromatography, Ion Exchange
  • DNA-Binding Proteins*
  • Electrophoresis, Polyacrylamide Gel
  • Molecular Sequence Data
  • Staphylococcus aureus / chemistry*
  • Staphylococcus aureus / physiology

Substances

  • Amino Acids
  • Bacterial Proteins
  • DNA-Binding Proteins
  • vitronectin-binding protein, Staphylococcus aureus