Detection of von Willebrand factor-cleaving protease (ADAMTS-13) in human platelets.
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.
Suzuki
Misako
M
Department of Internal Medicine, Keio University, Tokyo 160-8582, Japan.
Murata
Mitsuru
M
Matsubara
Yumiko
Y
Uchida
Toshihiro
T
Ishihara
Hiroaki
H
Shibano
Toshiro
T
Ashida
Shin-ichiro
S
Soejima
Kenji
K
Okada
Yasunori
Y
Ikeda
Yasuo
Y
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
D000199:Actins; D018951:Antigens, CD20; D012333:RNA, Messenger; D014841:von Willebrand Factor; D051722:ADAM Proteins; D008666:Metalloendopeptidases; D000071120:ADAMTS13 Protein; C099604:ADAMTS13 protein, human