Clumping factor A mediates binding of Staphylococcus aureus to human platelets.
The direct binding of bacteria to platelets may be an important virulence mechanism in the pathogenesis of infective endocarditis. We have previously described Staphylococcus aureus strain PS12, a Tn551-derived mutant of strain ISP479, with reduced ability to bind human platelets in vitro. When tested in an animal model of endocarditis, the PS12 strain was less virulent than its parental strain, as measured by bacterial densities in endocardial vegetations and incidence of systemic embolization. We have now characterized the gene disrupted in PS12 and its function in platelet binding. DNA sequencing, Southern blotting, and PCR analysis indicate that PS12 contained two Tn551 insertions within the clumping factor A (ClfA) locus (clfA). The first copy was upstream from the clfA start codon and appeared to have no effect on ClfA production. The second insertion was within the region encoding the serine aspartate repeat of ClfA and resulted in the production of a truncated ClfA protein that was secreted from the cell. A purified, recombinant form of the ClfA A region, encompassing amino acids 40 through 559, significantly reduced the binding of ISP479C to human platelets by 44% (P = 0.0001). Immunoprecipitation of recombinant ClfA that had been incubated with solubilized platelet membranes coprecipitated a 118-kDa platelet membrane protein. This protein does not appear to be glycoprotein IIb. These results indicate that platelet binding by S. aureus is mediated in part by the direct binding of ClfA to a novel 118-kDa platelet membrane receptor.
Siboo
I R
IR
Veterans Affairs Medical Center and University of California San Francisco, San Francisco, California, USA.
Cheung
A L
AL
Bayer
A S
AS
Sullam
P M
PM
D001422:Bacterial Adhesion; D001792:Blood Platelets / Q000382:microbiology*; D003030:Coagulase / Q000502:physiology*; D004251:DNA Transposable Elements; D005340:Fibrinogen / Q000502:physiology; D018628:Gene Dosage; D006801:Humans; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000032:analysis; D013211:Staphylococcus aureus / Q000502:physiology*; D014158:Transcription, Genetic
D003030:Coagulase; D004251:DNA Transposable Elements; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D005340:Fibrinogen