Cloning and expression of the gene for a fibronectin-binding protein from Staphylococcus aureus. Academic Article uri icon

abstract

  • The gene encoding the fibronectin-binding protein (FNBP) from Staphylococcus aureus strain 8325-4 was isolated from a gene bank in pBR322. The original clone, containing a 6.5-kb insert, gave a functional product present in the periplasm of Escherichia coli. Analysis of polypeptides isolated after affinity chromatography on fibronectin-Sepharose followed by ion-exchange chromatography revealed two gene products, 87 and 165 kd in mol. wt. The amino acid compositions of these two polypeptides and a native FNBP from S. aureus strain Newman were very similar. Antibodies raised against the native FNBP from strain Newman precipitated the 125I-labelled 165-kd polypeptide, and unlabeled 165- and 87-kd polypeptides as well as native FNBP inhibited the immunoprecipitation reactions. The region of the fnbp-gene encoding the fibronectin-binding activity has been identified and subcloned in an expression vector based on the staphylococcal protein A gene. The resulting product in E. coli is an extracellular fusion protein consisting of two IgG-binding domains of protein A followed by a fibronectin-binding region. The fusion protein binds to fibronectin and completely inhibits the binding of fibronectin to intact cells of S. aureus.

published proceedings

  • EMBO J

altmetric score

  • 9

author list (cited authors)

  • Flock, J. I., Frman, G., Jnsson, K., Guss, B., Signs, C., Nilsson, B., ... Lindberg, M.

citation count

  • 218

complete list of authors

  • Flock, JI||Fröman, G||Jönsson, K||Guss, B||Signäs, C||Nilsson, B||Raucci, G||Höök, M||Wadström, T||Lindberg, M

publication date

  • August 1987

publisher