The N-terminal of thrombospondin-1 is essential for coagulase-negative staphylococcal binding.
Bacterial binding was studied to determine whether thrombospondin-1 (TSP) acts as a ligand in attachment of coagulase-negative staphylococci (CNS). Twenty-five of 27 CNS strains bound soluble TSP. Staphylococcus epidermidis J9P bound 125I-labelled TSP in a dose-dependent manner. Scatchard plot analysis of the binding of TSP by strain J9P revealed two Kd values of 6.4 x 10(-9) M and 2.9 x 10(-8) M. The binding structures of strain J9P were sensitive to protease and were resistant to heat treatment. Unlabelled TSP and recombinant von Willebrand factor inhibited binding of TSP by strain J9P, but other proteins or monosaccharides did not. Heparin inhibited binding of TSP to strain J9P and two other S. epidermidis strains, BD5703 and BD969. Fusion proteins of the type 1 repeats, type 2 repeats, type 3 repeats and C-terminal domain of TSP or the synthetic Arg-Gly-Asp peptide did not inhibit binding of TSP to bacteria. TSP promoted adhesion of S. epidermidis strains when it was immobilised on polymer surfaces. These results indicate that the specific interaction between CNS and TSP may contribute to bacterial adhesion on biomaterial surfaces. The N-terminal heparin-binding domain of TSP appears to be the major region for recognition by CNS.
Yanagisawa
Naoko
N
Department of Medical Microbiology, Dermatology and Infection, Lund University, Lund, Sweden.
Li
Dai-Qing
DQ
Department of Medical Microbiology, Dermatology and Infection, Lund University, Lund, Sweden.
Ljungh
Åsa
Å
Department of Medical Microbiology, Dermatology and Infection, Lund University, Lund, Sweden.
D001422:Bacterial Adhesion / Q000502:physiology*; D001792:Blood Platelets; D003030:Coagulase; D003470:Culture Media; D004305:Dose-Response Relationship, Drug; D010450:Endopeptidases; D006358:Hot Temperature; D006801:Humans; D007456:Iodine Isotopes; D007700:Kinetics; D008024:Ligands; D012995:Solubility; D013212:Staphylococcus epidermidis / Q000201:enzymology / Q000378:metabolism*; D013329:Structure-Activity Relationship; D019700:Thrombospondin 1 / Q000737:chemistry / Q000378:metabolism*; D014841:von Willebrand Factor / Q000378:metabolism
D003030:Coagulase; D003470:Culture Media; D007456:Iodine Isotopes; D008024:Ligands; D019700:Thrombospondin 1; D014841:von Willebrand Factor; D010450:Endopeptidases