Crystallization and preliminary X-ray analysis of active site-inhibited human coagulation factor VIIa (des-Gla).
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 P41212 or P43212 with unit cell dimensions a = b = 94.85 A, c = 114.30 A, contain one molecule per asymmetric unit, and diffract to 2.3-A resolution when exposed to synchrotron radiation.
Johnson
D J
DJ
Haemostasis Research Group, MRC Clinical Sciences Centre, Imperial College School of Medicine, Du Cane Road, London, W12 0NN, United Kingdom.
Nugent
P G
PG
Tuddenham
E G
EG
Harlos
K
K
Kemball-Cook
G
G
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology*; D001665:Binding Sites; D003460:Crystallization; D018360:Crystallography, X-Ray; D003619:Dansyl Compounds / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015942:Factor VIIa / Q000037:antagonists & inhibitors / Q000737:chemistry*; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry; D017384:Sequence Deletion
D000590:Amino Acid Chloromethyl Ketones; D003619:Dansyl Compounds; D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; C030824:dansylglutamyl-glycyl-arginine chloromethyl ketone; D015942:Factor VIIa