Identification of aspartic acid 514 through glutamic acid 542 as a glycoprotein Ib-IX complex receptor recognition sequence in von Willebrand factor. Mechanism of modulation of von Willebrand factor by ristocetin and botrocetin.
As the first step in hemostasis, the binding of von Willebrand factor (vWF) to the platelet membrane glycoprotein (GP) Ib-IX complex is essential for platelet adhesion at high-shear blood flow. This interaction in vivo requires the prior binding of vWF to the subendothelial matrix, a process which exposes a normally cryptic binding site on vWF for the GP Ib-IX complex. This process can be mimicked in vitro by modulators such as ristocetin or the snake venom protein botrocetin or by desialation of vWF. We have previously localized the GP Ib binding site on vWF to a monomeric dispase fragment which extends from Leu-480/Val-481 to Gly-718 in the primary sequence of mature vWF [Andrews, R. K., Gorman, J. J., Booth, W. J., Corino, G. L., Castaldi, P. A., & Berndt, M. C. (1989) Biochemistry 28, 8326-8336]. This fragment also contains a distinct binding site for botrocetin. Analysis of synthetic peptides corresponding to hydrophilic stretches of sequence within this fragment indicated that the sequence Asp-514-Glu-542 represents a major adhesive sequence involved in receptor recognition. This peptide inhibited both the ristocetin- and botrocetin-mediated binding of vWF to either platelets or purified GP Ib-IX complex (IC50 approximately 50-200 microM) as well as the asialo-vWF- and bovine vWF-dependent agglutination of platelets. Both the N- and C-terminal halves of the peptide were inhibitory but less so than the intact peptide. This peptide also inhibited botrocetin binding to vWF, suggesting that botrocetin modulates vWF-GP Ib interaction by binding in close proximity to the vWF adhesion sequence.(ABSTRACT TRUNCATED AT 250 WORDS)
Berndt
M C
MC
Vascular Biology Laboratory, Baker Medical Research Institute, Prahran, Victoria, Australia.
Ward
C M
CM
Booth
W J
WJ
Castaldi
P A
PA
Mazurov
A V
AV
Andrews
R K
RK
D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites; D003435:Crotalid Venoms / Q000494:pharmacology*; D005971:Glutamates / Q000235:genetics* / Q000378:metabolism; D018698:Glutamic Acid; D006388:Hemagglutinins / Q000494:pharmacology; D007457:Iodine Radioisotopes; D008969:Molecular Sequence Data; D010980:Platelet Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D012310:Ristocetin / Q000494:pharmacology*; D013056:Spectrophotometry, Ultraviolet; D014841:von Willebrand Factor / Q000378:metabolism*
D003435:Crotalid Venoms; D005971:Glutamates; D006388:Hemagglutinins; D007457:Iodine Radioisotopes; D010980:Platelet Membrane Glycoproteins; D014841:von Willebrand Factor; D012310:Ristocetin; D001224:Aspartic Acid; D018698:Glutamic Acid; C030961:botrocetin