Abstract
Endothelin (ET) is a 21-residue potent vasoconstrictive peptide produced by vascular endothelial cells and formed from its precursor, big endothelin (big ET), by endothelin-converting enzyme (ECE). This paper describes the cloning and functional expression of a cDNA encoding a human ECE from human umbilical vein endothelial cells (HUVEC). Human ECE consists of 758 amino acid residues and has high homology to rat and bovine ECE. Immunoblot analysis using a monoclonal antibody risen against rat lung ECE showed the presence of immunoreactive protein in membrane fraction prepared from both HUVEC and COS-1 cells transfected with human ECE cDNA. Both COS-1 cells expressing human ECE and its membrane fraction converted big ET-1 most efficiently among big ETs.
MeSH terms
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Amino Acid Sequence
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Animals
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Aspartic Acid Endopeptidases / biosynthesis*
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Aspartic Acid Endopeptidases / isolation & purification
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Aspartic Acid Endopeptidases / metabolism
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Base Sequence
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Cattle
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Cell Line
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Cells, Cultured
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Chlorocebus aethiops
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Cloning, Molecular
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Conserved Sequence
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DNA, Complementary / metabolism
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Endothelin-1
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Endothelin-Converting Enzymes
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Endothelins / metabolism
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Endothelium, Vascular / enzymology*
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Gene Library
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Humans
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Immunoblotting
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Kidney
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Metalloendopeptidases
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Molecular Sequence Data
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Protein Precursors / metabolism
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Rats
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Sequence Homology, Amino Acid
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Transfection
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Umbilical Veins
Substances
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DNA, Complementary
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Endothelin-1
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Endothelins
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Protein Precursors
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Recombinant Proteins
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Aspartic Acid Endopeptidases
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Metalloendopeptidases
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Endothelin-Converting Enzymes