Abstract
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.
MeSH terms
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ADAM Proteins
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ADAMTS13 Protein
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Amino Acid Sequence
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Amino Acids / chemistry
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Animals
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Blotting, Northern
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Blotting, Western
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Cell Line
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Cloning, Molecular
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DNA Primers / chemistry
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DNA, Complementary / metabolism
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Databases, Genetic
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Dose-Response Relationship, Drug
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Edetic Acid / pharmacology
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Humans
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Metalloendopeptidases / chemistry
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Metalloendopeptidases / genetics*
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Metalloendopeptidases / metabolism*
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Mice
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Mice, Inbred BALB C
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Models, Genetic
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Molecular Sequence Data
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Phylogeny
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Poly A / metabolism
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Polymerase Chain Reaction
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Protein Binding
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Protein Structure, Tertiary
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Purpura, Thrombotic Thrombocytopenic / blood
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RNA / metabolism
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RNA, Messenger / metabolism
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Recombinant Proteins / chemistry
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Sequence Homology, Amino Acid
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Tissue Distribution
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Transfection
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von Willebrand Factor / metabolism
Substances
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Amino Acids
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DNA Primers
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DNA, Complementary
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RNA, Messenger
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Recombinant Proteins
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von Willebrand Factor
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Poly A
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RNA
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Edetic Acid
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ADAM Proteins
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ADAMTS13 protein, mouse
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Metalloendopeptidases
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ADAMTS13 Protein
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ADAMTS13 protein, human