Isolation and characterization of collagen-binding domains from human von Willebrand factor |
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| Authors: | N E Sharapova A P Kotnova Z M Galushkina N N Poletaeva N V Lavrova E I Aksenova A S Semikhin A S Karyagina and V G Lunin |
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| Institution: | (1) Department of Chemistry and Biochemistry, University of Arkansas, Fayetteville, AR 72701, USA;(2) Department of Microbiology & Parasitology, Kitasato University Medical School, 1-15-1 Kitasato, Sagamihara Kanagawa, 228-8555, Japan;(3) Present address: Department of Molecular Structure, Amgen, Inc., MS 14-2-A, One Amgen Center Drive, Thousand Oaks, CA 91320, USA |
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| Abstract: | DNA fragments that encode two collagen-binding decapeptides from human von Willebrand factors vWF-H1 and vWF-H2 were cloned
in Escherichia coli cells. The effective chimeric proteins vWF(H1)-CBD and vWF(H2)-CBD, which produce strains that contain corresponding decapeptide
sequences, Gly-Ser spacer, and a cellulose-binding domain (CBD) from Anaerocellum thermophilum were constructed. Highly purified samples of vWF(H1)-CBD and vWF(H2)-CBD proteins were obtained using the one-stage purification
method on cellulose and their ability to bind collagen was studied. The obtained constructions are planned to for use in the
development of recombinant collagen-binding proteins with different biological activity, which will be used for the further
development of a new generation of products and materials for medical purposes, e.g., various kids of implants and coatings. |
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