GST-Fbe can recognize beta-chains of fibrin(ogen) on explanted materials.
Staphylococcus epidermidis, a coagulase-negative staphylococcus (CoNS), is one of the leading pathogens of nosocomial infections, particularly associated with foreign body infections. Adherence of S. epidermidis to fibrinogen deposited on the surfaces of implants is important for the development of foreign body infections. A gene (fbe) encoding a fibrinogen-binding protein from S. epidermidis (Fbe) was identified by shotgun phage display. A portion of fbe was cloned into a GST-fusion vector. Affinity to glutathione-Sepharose by the GST-tag and affinity to fibrinogen-Sepharose by the Fbe part were applied to purify the recombinant Fbe. The purity and efficacy of the methods used in protein purification was compared. Furthermore, the potential physiological role of Fbe was studied by the interaction between GST-Fbe and components extracted from explanted materials in vitro.
Pei
Lei
L
Division of Clinical Bacteriology, Department of Microbiology, Pathology and Immunology, Huddinge University Hospital, Karolinska Institute, SE-141 86, Stockholm, Sweden. lpei@parners.org
Arvholm
Ingegerd Löfving
IL
Lonnies
Lena
L
Flock
Jan-Ingmar
JI
D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002846:Chromatography, Affinity; D005340:Fibrinogen / Q000378:metabolism*; D005982:Glutathione Transferase / Q000737:chemistry / Q000378:metabolism*; D008567:Membranes, Artificial; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism*
D001426:Bacterial Proteins; D002352:Carrier Proteins; C112556:Fbe protein, bacteria; D008567:Membranes, Artificial; D011993:Recombinant Fusion Proteins; D005340:Fibrinogen; D005982:Glutathione Transferase