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Cloning, expression and functional characterization of the full‐length murine ADAMTS13
Author(s) -
BRUNO K.,
VÖLKEL D.,
PLAIMAUER B.,
ANTOINE G.,
PABLE S.,
MOTTO D. G.,
LEMMERHIRT H. L.,
DORNER F.,
ZIMMERMANN K.,
SCHEIFLINGER F.
Publication year - 2005
Publication title -
journal of thrombosis and haemostasis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.947
H-Index - 178
eISSN - 1538-7836
pISSN - 1538-7933
DOI - 10.1111/j.1538-7836.2005.01246.x
Subject(s) - cloning (programming) , computational biology , expression cloning , microbiology and biotechnology , biology , chemistry , genetics , computer science , gene , complementary dna , programming language
Summary. Functional deficiency or absence of the human von Willebrand factor (VWF)‐cleaving protease (VWF‐cp), recently termed ADAMTS13, has been shown to cause acquired and congenital thrombotic thrombocytopenic purpura (TTP), respectively. As a first step towards developing a small animal model of TTP, we have cloned the complete (non‐truncated) murine Adamts13 gene from BALB/c mice liver poly A + mRNA. Murine ADAMTS13 is a 1426‐amino‐acid protein with a high homology and similar structural organization to the human ortholog. Transient expression of the murine Adamts13 cDNA in HEK 293 cells yielded a protein with a molecular weight of approximately 180 kDa which degraded recombinant murine VWF (rVWF) in a dose‐dependent manner. The cleavage products of murine rVWF had the expected size of 140 and 170 kDa. Murine ADAMTS13 was inhibited by EDTA and the plasma from a TTP patient.