Allosteric activation of coagulation factor VIIa visualized by hydrogen exchange.

Full Abstract

Coagulation factor VIIa (FVIIa) is a serine protease that, after binding to tissue factor (TF), plays a pivotal role in the initiation of blood coagulation. We used hydrogen exchange monitored by mass spectrometry to visualize the details of FVIIa activation by comparing the exchange kinetics of distinct molecular states, namely zymogen FVII, endoproteolytically cleaved FVIIa, TF-bound zymogen FVII, TF-bound FVIIa, and FVIIa in complex with an active site inhibitor. The hydrogen exchange kinetics of zymogen FVII and FVIIa are identical indicating highly similar solution structures. However, upon tissue factor binding, FVIIa undergoes dramatic structural stabilization as indicated by decreased exchange rates localized throughout the protease domain and in distant parts of the light chain, spanning across 50A and revealing a concerted interplay between functional sites in FVIIa. The results provide novel insights into the cofactor-induced activation of this important protease and reveal the potential for allosteric regulation in the trypsin family of proteases.

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Author information

Author/s: Rand, Kasper D (KD); Jørgensen, Thomas J D (TJ); Olsen, Ole H (OH); Persson, Egon (E); Jensen, Ole N (ON); Stennicke, Henning R (HR); Andersen, Mette D (MD);

Affiliation: Department of Haemostasis Biochemistry, Novo Nordisk A/S, Novo Nordisk Park, DK-2760 Måløv, Denmark. kprn(-atsign-)novonordisk.com

Journal and publication information

Publication Type: Journal Article; Research Support, Non-U.S. Gov't

Journal: The Journal of biological chemistry (J Biol Chem), published in United States. (Language: eng)

Reference: 2006-Aug; vol 281 (issue 32) : pp 23018-24

Dates: Created 2006/08/07; Completed 2006/11/08; Revised 2006/11/15;

PMID: 16687401, status: MEDLINE (last retrieval date: 12/26/2008)

Sourced from the National Library of Medicine. Abstract text and other information may be subject to copyright.

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MeSH headings (categories)

This article was linked to the MESH Headings shown below.

Allosteric Site
Enzyme Activation
Enzyme Precursors - chemistry
Factor VIIa - chemistry; metabolism
Humans
Hydrogen - chemistry
Kinetics

Mass Spectrometry - methods
Models, Molecular
Molecular Conformation
Peptide Hydrolases - chemistry
Protein Structure, Tertiary
Recombinant Proteins - chemistry
Trypsin - chemistry

Associated Chemicals: Enzyme Precursors (0) ; Recombinant Proteins (0) ; Hydrogen (1333-74-0) ; Peptide Hydrolases (EC 3.4.-) ; Factor VIIa (EC 3.4.21.21) ; Trypsin (EC 3.4.21.4)

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