Human recombinant factor XIII from Saccharomyces cerevisiae. Crystallization and preliminary x-ray data.

P.D Bishop,G.W Lasser,I Le Trong,R.E Stenkamp,D.C Teller

doi:10.1016/s0021-9258(18)77431-0

Abstract

Crystals of human recombinant factor XIII from the yeast Saccharomyces cerevisiae have been grown from solutions of ammonium sulfate at pH 5.8. The crystals are orthorhombic, with space group P2(1)2(1)2 and unit cell dimensions gamma a = 101.2, b = 182.7, and c = 93.4 A. The asymmetric unit consists of one a2 dimer of molecular mass 166 kDa. A 3.5-A resolution data set for the native protein has been collected. Practical resolution limits for these crystals have not been determined, but reflections have been observed to a Bragg spacing of 2.8-A resolution.

Highlights

Crystals of human recombinant factor XIII from the yeast Saccharomyces cerevisiae have been grown from solutions of ammonium sulfate at pH 5.8
Factor XIII is the last enzyme to become activated in the blood coagulation cascade and is the focus of therapeutic strategies aimed at controlling various clotting abnormalities and tissue injuries
The activation of factor XIII is a highly regulated process, with fibrin serving as a cofactor both in the calcium-dependent conformational change necessary for exposure of the active-site thiol [15, 16] and in the initial proteolytic event

Summary

From the Department of Biochemistry

Crystals of human recombinant factor XIII from the yeast Saccharomyces cerevisiae have been grown from solutions of ammonium sulfate at pH 5.8. Factor XIII is the last enzyme to become activated in the blood coagulation cascade and is the focus of therapeutic strategies aimed at controlling various clotting abnormalities and tissue injuries It functions as a transglutaminase which catalyzes the formation of y-glutamyl-t-lysylamide crosslinks between polypeptide chains in adjacent fibrin monomers and between fibrin and other plasma proteins (l-4). DNA sequence analysis revealed that separate exons encode the activation peptide released by thrombin, the active-site cysteine region, the two putative calcium-binding regions, and the thrombin cleavage site leading to inactivation. This suggeststhat the introns may separate the a subunit into functional and structural domains [21]. We report here the crystallization and preliminary diffraction results for this recombinant factor XIII

AND METHODS

The solution was incubated on ice

RESULTS AND DISCUSSION