Identification of a binding site for glycoprotein Ibalpha in the Apple 3 domain of factor XI.
Factor XI (FXI) is a homodimeric plasma zymogen that is cleaved at two internal Arg(369)-Ile(370) bonds by thrombin, factor XIIa, or factor XIa. FXI circulates as a complex with the glycoprotein high molecular weight kininogen (HK). FXI binds to specific sites (K(d) = approximately 10 nM, B(max) = approximately 1,500/platelet) on the surface of stimulated platelets, where it is efficiently activated by thrombin. The FXI Apple 3 (A3) domain mediates binding to platelets in the presence of HK and zinc ions (Zn(2+)) or prothrombin and calcium ions. The platelet glycoprotein (GP) Ib-IX-V complex is the receptor for FXI. Using surface plasmon resonance, we determined that FXI binds specifically to glycocalicin, the extracellular domain of GPIbalpha, in a Zn(2+)-dependent fashion (K(d) = approximately 52 nM). We now show that recombinant FXI A3 domain inhibits FXI inbinding to glycocalicin in the presence of Zn(2+), whereas the recombinant FXI A1, A2, or A4 domains have no effect. Experiments with full-length recombinant FXI mutants show that, in the presence of Zn(2+), glycocalicin binds FXI at a heparin-binding site in A3 (Lys(252) and Lys(253)) and not by amino acids previously shown to be required for platelet binding (Ser(248), Arg(250), Lys(255), Phe(260), and Gln(263)). However, binding in the presence of HK and Zn(2+) requires Ser(248), Arg(250), Lys(255), Phe(260), and GLn(263) and not Lys(252) and Lys(253). Thus, binding of FXI to GPIbalpha is mediated by amino acids in the A3 domain in the presence or absence of HK. This interaction is important for the initiation of the consolidation phase of blood coagulation and the generation of thrombin at sites of platelet thrombus formation.
Sol Sherry Thrombosis Research Center, Temple University School oof Medicine, Philadelphia, PA 19140, USA.
Frank A
FA
Baglia
David
D
Gailani
José A
JA
López
Peter N
PN
Walsh
D001665:Binding Sites; D001792:Blood Platelets / Q000378:metabolism; D002712:Chlorides / Q000494:pharmacology; D005172:Factor XI / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D019679:Kininogen, High-Molecular-Weight / Q000494:pharmacology; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000378:metabolism*; D019038:Platelet Glycoprotein GPIb-IX Complex / Q000378:metabolism; D017967:Zinc Compounds / Q000494:pharmacology
D002712:Chlorides; D019679:Kininogen, High-Molecular-Weight; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D019038:Platelet Glycoprotein GPIb-IX Complex; D017967:Zinc Compounds; C094990:adhesion receptor; C023592:glycocalicin; C016837:zinc chloride; D005172:Factor XI