Thrombospondin binds to Staphylococcus aureus and promotes staphylococcal adherence to surfaces.
Bacterial adherence to surfaces is the determining first step in staphylococcal infections. Activated platelets mediate adherence of staphylococci to tissues during inflammation or infection; however, the molecular mechanisms of this interaction are not clearly understood. Thrombospondin, a large multifunctional glycoprotein, is the principal platelet-stored glycoprotein. It is secreted upon platelet activation and either bound to receptors on the platelet surface or released and incorporated into blood clots and extracellular matrices. To characterize thrombospondin binding to staphylococci, we incubated [125I]thrombospondin with Staphylococcus aureus Cowan 1 in the presence of albumin and separated bound and free thrombospondin by centrifugation. We found that binding was (i) specific, since it was up to 76% inhibitable and up to 60% reversible in the presence of a 100-fold excess of unlabeled thrombospondin, (ii) saturable, with an apparent dissociation constant (Kd) of 5.6 x 10(-9) M and a maximal number of 2,600 binding sites per microorganism, and (iii) Ca2+ dependent, since omission of this ion from the medium decreased significantly the binding capacity. The binding reaction was insensitive to previous trypsin treatment of bacteria, but it was strongly inhibited in the presence of heparin. Protein A-negative and -positive strains had similar binding characteristics. To determine the promotion of staphylococcal adherence to surfaces by solid-phase thrombospondin, we incubated 3H-labeled S. aureus Cowan 1 and 26 pathogenic staphylococcal isolates with thrombospondin-coated polymethylmethacrylate disks and found that adherence was significantly promoted as a function of adsorbed thrombospondin. These results indicate a role for thrombospondin as an important mediator of staphylococcal adherence to activated platelets, to blood clots, or to extracellular matrices in pyogenic infections.
Herrmann
M
M
Department of Medicine, University Hospital, Geneva, Switzerland.
Suchard
S J
SJ
Boxer
L A
LA
Waldvogel
F A
FA
Lew
P D
PD
D001422:Bacterial Adhesion; D001665:Binding Sites; D001792:Blood Platelets / Q000502:physiology*; D002118:Calcium / Q000502:physiology; D005340:Fibrinogen / Q000494:pharmacology; D005353:Fibronectins / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000502:physiology*; D019699:Thrombospondins
D005353:Fibronectins; D010980:Platelet Membrane Glycoproteins; D019699:Thrombospondins; D005340:Fibrinogen; D006493:Heparin; D002118:Calcium