Document Detail


N-terminal flanking region of A1 domain in von Willebrand factor stabilizes structure of A1A2A3 complex and modulates platelet activation under shear stress.
MedLine Citation:
PMID:  22431729     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
von Willebrand factor (vWF) mediates platelet adhesion and thrombus formation via its interaction with the platelet receptor glycoprotein (GP)Ibα. We have analyzed two A1A2A3 tri-domain proteins to demonstrate that the amino acid sequence, Gln(1238)-Glu(1260), in the N-terminal flanking region of the A1 domain, together with the association between the A domains, modulates vWF-GPIbα binding and platelet activation under shear stress. Using circular dichroism spectroscopy and differential scanning calorimetry, we have described that sequence Gln(1238)-Glu(1260) stabilizes the structural conformation of the A1A2A3 tri-domain complex. The structural stabilization imparted by this particular region inhibits the binding capacity of the tri-domain protein for GPIbα. Deletion of this region causes a conformational change in the A1 domain that increases binding to GPIbα. Only the truncated protein was capable of effectively blocking ristocetin-induced platelet agglutination. To determine the capacity of activating platelets via the interaction with GPIbα, whole blood was incubated with the N-terminal region truncated or intact tri-A domain protein prior to perfusion over a fibrin(ogen)-coated surface. At a high shear rate of 1,500 s(-1), platelets from blood containing the truncated protein rapidly bound, covering >90% of the fibrin(ogen) surface area, whereas the intact tri-A domain protein induced platelets to bind <10%. The results obtained in this study ascertain the relevant role of the structural association between the N-terminal flanking region of the A1 domain (amino acids Gln(1238)-Glu(1260)) and the A1A2A3 domain complex in preventing vWF to bind spontaneously to GPIbα in solution under high shear forces.
Authors:
Matthew Auton; Katie E Sowa; Molly Behymer; Miguel A Cruz
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Publication Detail:
Type:  Clinical Trial; Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2012-03-19
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  287     ISSN:  1083-351X     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2012 Apr 
Date Detail:
Created Date:  2012-04-30     Completed Date:  2012-07-02     Revised Date:  2013-05-20    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  14579-85     Citation Subset:  IM    
Affiliation:
Section of Cardiovascular Research, Department of Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.
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MeSH Terms
Descriptor/Qualifier:
Anti-Bacterial Agents / pharmacology
Female
Fibrinogen / genetics,  metabolism
Humans
Male
Membrane Glycoproteins / genetics,  metabolism*
Multiprotein Complexes / genetics,  metabolism
Platelet Activation / drug effects,  physiology*
Protein Structure, Quaternary
Protein Structure, Tertiary
Ristocetin / pharmacology
Stress, Physiological / drug effects,  physiology*
von Willebrand Factor / genetics,  metabolism*
Grant Support
ID/Acronym/Agency:
HL109109/HL/NHLBI NIH HHS; HL72886/HL/NHLBI NIH HHS; R01 HL109109/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Anti-Bacterial Agents; 0/Membrane Glycoproteins; 0/Multiprotein Complexes; 0/adhesion receptor; 0/von Willebrand Factor; 1404-55-3/Ristocetin; 9001-32-5/Fibrinogen
Comments/Corrections

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