Fibrinogen-binding protein/clumping factor from Staphylococcus aureus.
The binding of staphylococcal components to fibrinogen was studied. Fibrinogen-binding material from lysed staphylococcal cells or culture supernatants was affinity purified on fibrinogen-Sepharose and analyzed on Western (immuno-) blots by the use of fibrinogen and antifibrinogen antibodies. Two main bands of 87 and 19 kilodaltons (kDa) and a weaker band of 35 kDa bound specifically to fibrinogen. A monoclonal antibody bound to all three bands, indicating that these were of the same origin. The yield of these components was much higher in the culture supernatant than on washed cells, suggesting that these molecules are essentially extracellular products. In a plasma coagulase test, the 87-kDa band, but not the 19-kDa band, clotted rabbit plasma, demonstrating that the 87-kDa molecule is coagulase. This was further confirmed by the fact that the 87-kDa band binds specifically to prothrombin. It was shown that the 87- and the 19-kDa molecules were present on the cell surface by surface labeling the cells with 125I. In addition, the fact that killed and washed cells could induce plasma clotting demonstrates that staphylococci have coagulase exposed on the surface. It was concluded that cell-bound coagulase has affinity for fibrinogen also in the absence of prothrombin and thus is responsible for the clumping of staphylococci in fibrinogen.
Bodén
M K
MK
Center for Biotechnology, Karolinska Institute, Huddinge, Sweden.
Flock
J I
JI
D001426:Bacterial Proteins / Q000097:blood / Q000302:isolation & purification*; D015153:Blotting, Western; D002352:Carrier Proteins / Q000097:blood / Q000302:isolation & purification*; D002458:Cell Fractionation; D002846:Chromatography, Affinity; D003030:Coagulase / Q000097:blood / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000378:metabolism*; D008970:Molecular Weight; D012967:Sodium Dodecyl Sulfate; D013211:Staphylococcus aureus / Q000032:analysis* / Q000254:growth & development
D001426:Bacterial Proteins; D002352:Carrier Proteins; D003030:Coagulase; D012967:Sodium Dodecyl Sulfate; D005340:Fibrinogen