Crystallization and preliminary X-ray analysis of active site-inhibited human coagulation factor VIIa (des-Gla)

被引:6
作者
Johnson, DJD
Nugent, PG
Tuddenham, EGD
Harlos, K
Kemball-Cook, G
机构
[1] Hammersmith Hosp, Imperial Coll Sch Med, MRC, Clin Sci Ctr,Haemostasis Res Grp, London W12 0NN, England
[2] Oxford Ctr Mol Sci, Lab Mol Biophys, Oxford OX1 3QU, England
关键词
coagulation; crystallization; factor VII; haemostasis; protein purification; recombinant expression; X-ray diffraction;
D O I
10.1006/jsbi.1998.4078
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Human coagulation factor VIIai that lacks the Gla domain (residues 1-44) has been prepared, purified, and crystallised, First, recombinant factor VII was activated to form factor VIIa, the active site was then inhibited with 1,5-dansyl-Glu-Gly-Arg-chloromethyl ketone, and finally the Gla domain was removed by chymotryptic digestion, yielding factor VIIai (des-Gla). After further purification single crystals suitable for x-ray analysis were obtained by vapour diffusion. Crystals of factor VIIai (des-Gla) belong to the tetragonal space group P4(1)2(1)2 or P4(3)2(1)2 with unit cell dimensions a = b = 94.85 Angstrom, c = 114.30 Angstrom contain one molecule per asymmetric unit, and diffract to 2.3-Angstrom resolution when exposed to synchrotron radiation. (C) 1999 Academic Press.
引用
收藏
页码:90 / 93
页数:4
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