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Title: Solution structure of human von Willebrand factor studied using small angle nuetron scattering

Abstract

Von Willebrand factor (vWF) binding to platelets under high fluid shear is an important step regulating atherothrombosis. We applied light and small-angle neutron scattering to study the solution structure of human vWF multimers and protomer. Results suggest that these proteins resemble prolate ellipsoids with radius of gyration (Rg) of ~75nm and ~30nm for multimer and protomer respectively. The ellipsoid dimensions/radii are 175×28nm for multimers and 70×9.1nm for protomers. Substructural repeat domains are evident within multimeric-vWF that are indicative of elements of the protomer quarternary structure (16nm) and individual functional domains (4.5nm). Amino acids occupy only ~2% volume of multimer and protomer ellipsoids, compared to other proteins like albumin (98%) and fibrinogen (35%). vWF treatment with Guanidine·HCl, which increases vWF susceptibility to proteolysis by ADAMTS-13, causes local structural changes at length scales<10nm without altering protein Rg. Treatment of multimer but not protomer-vWF with random homobifunctional linker BS3, prior to reduction of inter-monomer disulfide linkages and western blotting reveals a pattern of dimer and trimer units that indicate the presence of stable inter-monomer non-covalent interactions within the multimer. Overall, vWF solution structure is stabilized by non-covalent interactions between different monomer units. Local changes in multimer conformation are sufficient for ADAMTS-13 mediated proteolysis.

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
918859
Report Number(s):
PNNL-SA-49749
Journal ID: ISSN 0021-9258; JBCHA3; TRN: US0805917
DOE Contract Number:  
AC05-76RL01830
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Biological Chemistry; Journal Volume: 281; Journal Issue: 50
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS; ALBUMINS; AMINO ACIDS; DIMERS; DISULFIDES; FIBRINOGEN; FUNCTIONALS; GUANIDINES; MONOMERS; NEUTRONS; PROTEINS; PROTEOLYSIS; SCATTERING; SHEAR

Citation Formats

Singh, Indrajeet, Shankaran, Harish, Beauharnois, Mark E, Xiao, Zhihua, Alexandridis, Paschalis, and Neelamegham, Sriram. Solution structure of human von Willebrand factor studied using small angle nuetron scattering. United States: N. p., 2006. Web. doi:10.1074/jbc.M607123200.
Singh, Indrajeet, Shankaran, Harish, Beauharnois, Mark E, Xiao, Zhihua, Alexandridis, Paschalis, & Neelamegham, Sriram. Solution structure of human von Willebrand factor studied using small angle nuetron scattering. United States. doi:10.1074/jbc.M607123200.
Singh, Indrajeet, Shankaran, Harish, Beauharnois, Mark E, Xiao, Zhihua, Alexandridis, Paschalis, and Neelamegham, Sriram. Fri . "Solution structure of human von Willebrand factor studied using small angle nuetron scattering". United States. doi:10.1074/jbc.M607123200.
@article{osti_918859,
title = {Solution structure of human von Willebrand factor studied using small angle nuetron scattering},
author = {Singh, Indrajeet and Shankaran, Harish and Beauharnois, Mark E and Xiao, Zhihua and Alexandridis, Paschalis and Neelamegham, Sriram},
abstractNote = {Von Willebrand factor (vWF) binding to platelets under high fluid shear is an important step regulating atherothrombosis. We applied light and small-angle neutron scattering to study the solution structure of human vWF multimers and protomer. Results suggest that these proteins resemble prolate ellipsoids with radius of gyration (Rg) of ~75nm and ~30nm for multimer and protomer respectively. The ellipsoid dimensions/radii are 175×28nm for multimers and 70×9.1nm for protomers. Substructural repeat domains are evident within multimeric-vWF that are indicative of elements of the protomer quarternary structure (16nm) and individual functional domains (4.5nm). Amino acids occupy only ~2% volume of multimer and protomer ellipsoids, compared to other proteins like albumin (98%) and fibrinogen (35%). vWF treatment with Guanidine·HCl, which increases vWF susceptibility to proteolysis by ADAMTS-13, causes local structural changes at length scales<10nm without altering protein Rg. Treatment of multimer but not protomer-vWF with random homobifunctional linker BS3, prior to reduction of inter-monomer disulfide linkages and western blotting reveals a pattern of dimer and trimer units that indicate the presence of stable inter-monomer non-covalent interactions within the multimer. Overall, vWF solution structure is stabilized by non-covalent interactions between different monomer units. Local changes in multimer conformation are sufficient for ADAMTS-13 mediated proteolysis.},
doi = {10.1074/jbc.M607123200},
journal = {Journal of Biological Chemistry},
number = 50,
volume = 281,
place = {United States},
year = {Fri Dec 15 00:00:00 EST 2006},
month = {Fri Dec 15 00:00:00 EST 2006}
}