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J. Biol. Chem., Vol. 266, Issue 13, 8149-8155, May, 1991
JP Scott, RR Montgomery and GS Retzinger
Ristocetin in aqueous solution dimerizes with an equilibrium dissociation
constant of 5.0 x 10(-4) M, i.e. approximately 1.1 mg ml-1 (Waltho, J.P.,
and Williams, D. H. (1989) J. Am. Chem. Soc. 111, 2475- 2480). At
concentrations of about 1.0 mg ml-1 ristocetin flocculates many proteins,
lyses platelets and, in the presence of von Willebrand factor, agglutinates
both fresh and formalin-fixed platelets. Because ristocetin exists as both
monomeric and dimeric species, we sought to determine which of these forms
flocculates proteins and agglutinates platelets. We found that: 1) the
initial rate of flocculation of certain proteins, 2) the initial rate of
agglutination of formalin- fixed platelets, and 3) the binding of
ristocetin to formalin-fixed platelets are higher order solely with respect
to the concentration of ristocetin dimers. As to the operative mechanism,
it appears that bifunctional dimers cross-link proteins that possess
multiple copies of a common recognition site. Preliminary evidence
indicates that a recognition site is a beta-turn of the form X-P-G-X'.
Dimeric ristocetin flocculates proteins, binds to platelets, and mediates von Willebrand factor-dependent agglutination of platelets
Department of Pediatrics, Medical College of Wisconsin, Milwaukee 53226.
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