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...

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Bibliographic Details
Published in:Journal of structural biology 1999-03, Vol.125 (1), p.90-93
Main Authors: Johnson, Daniel J.D., Nugent, Philip G., Tuddenham, Edward G.D., Harlos, Karl, Kemball-Cook, Geoffrey
Format: Article
Language:English
Subjects:
Amino Acid Chloromethyl Ketones - pharmacology
Binding Sites
coagulation
Crystallization
Crystallography, X-Ray
Dansyl Compounds - pharmacology
Enzyme Inhibitors - pharmacology
factor VII
Factor VIIa - antagonists & inhibitors
Factor VIIa - chemistry
haemostasis
Humans
protein purification
recombinant expression
Recombinant Proteins - chemistry
Sequence Deletion
X-ray diffraction
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Description
Summary: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 groupP41212 orP43212 with unit cell dimensionsa=b= 94.85 Å,c= 114.30 Å, contain one molecule per asymmetric unit, and diffract to 2.3-Å resolution when exposed to synchrotron radiation.
ISSN:1047-8477
1095-8657
DOI:10.1006/jsbi.1998.4078