Crystallization and preliminary X-ray analysis of coagulation factor IX-binding protein from habu snake venom at pH...

Coagulation factor IX-binding protein isolated from Trimeresurus flavoviridis (IX-bp) is a C-type lectin-like protein. It is an anticoagulant protein consisting of homologous subunits A and B. The subunits both contain a Ca2+-binding site with differing affinity (Kd values of 14 and 130 microM at pH 7.5). These binding characteristics are pH-dependent; under acidic conditions, the affinity of the low-affinity site was reduced considerably. In order to identify which site has high affinity and also to investigate the Ca2+-releasing mechanism, IX-bp was crystallized at pH 6.5 and 4.6. The crystals at pH 6.5 and 4.6 diffracted to 1.72 and 2.29 A resolution, respectively; the former crystals belong to the monoclinic space group P2(1), with unit-cell parameters a = 60.7, b = 63.5, c = 66.9 A, beta = 117.0 degrees, while the latter belong to the monoclinic space group C2, with a = 134.1, b = 37.8, c = 55.8 A, beta = 110.4 degrees.