title: Detection of von Willebrand factor-cleaving protease (ADAMTS-13) in human platelets.
abstract: The hemostatic activity of von Willebrand factor (vWF) is strongly dependent on its multimeric structure, with the highest activity in 'unusually large' multimers secreted from endothelial cells. The multimeric structure is regulated by a plasma protease, vWF-cleaving protease (vWF-CP, or ADAMTS-13). ADAMTS-13 mRNA is variably expressed in liver and other tissues. Because 15-25% of circulating vWF is stored in platelets, the presence and function of ADAMTS-13 in platelets are important issues. Here we report ADAMTS-13 expression in human platelets. Western blot analysis and flow cytometric analysis on permeabilized platelets revealed the presence of ADAMTS-13 protein. Real-time PCR demonstrated that ADAMTS-13 mRNA is present in platelets of six healthy volunteers, with little quantitative difference. The presence of ADAMTS-13 in platelets may imply the functional role of this enzyme in the local regulation of platelet function at the site of vascular injury or thrombus formation, and provide a useful tool for the analysis of structure and function of this enzyme.
authors.affiliation: Department of Internal Medicine, Keio University, Tokyo 160-8582, Japan.
authors.forename: Misako
authors.initials: M
authors.lastname: Suzuki
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Murata
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Matsubara
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Uchida
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Ishihara
authors.forename: Toshiro
authors.initials: T
authors.lastname: Shibano
authors.forename: Shin-ichiro
authors.initials: S
authors.lastname: Ashida
authors.forename: Kenji
authors.initials: K
authors.lastname: Soejima
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Okada
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Ikeda
mesh_terms: D051722:ADAM Proteins; D000071120:ADAMTS13 Protein; D000199:Actins / Q000032:analysis / Q000096:biosynthesis; D018951:Antigens, CD20 / Q000032:analysis / Q000096:biosynthesis; D001792:Blood Platelets / Q000201:enzymology*; D015153:Blotting, Western; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000097:blood*; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D014841:von Willebrand Factor / Q000378:metabolism
chemical_list: D000199:Actins; D018951:Antigens, CD20; D012333:RNA, Messenger; D014841:von Willebrand Factor; D051722:ADAM Proteins; D008666:Metalloendopeptidases; D000071120:ADAMTS13 Protein; C099604:ADAMTS13 protein, human