Molecular characterization of human and bovine endothelin converting enzyme (ECE-1).
A membrane-bound protease activity that specifically converts Big endothelin-1 has been purified from bovine endothelial cells (FBHE). The enzyme was cleaved with trypsin and the peptide sequencing analysis confirmed it to be a zinc chelating metalloprotease containing the typical HEXXH (HELTH) motif. RT-PCR and cDNA screens were employed to isolate the complete cDNAs of the bovine and human enzymes. This human metalloprotease was expressed heterologously in cell culture and oocytes. The catalytic activity of the recombinant enzyme is the same as that determined for the natural enzyme. The data suggest that the characterized enzyme represents the functional human endothelin converting enzyme ECE-1.
Schmidt
M
M
Department of Pharmaceutical Research, BASF Aktiengesellschaft, Ludwigshafen, Germany.
Kröger
B
B
Jacob
E
E
Seulberger
H
H
Subkowski
T
T
Otter
R
R
Meyer
T
T
Schmalzing
G
G
Hillen
H
H
D000595:Amino Acid Sequence; D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D015152:Blotting, Northern; D002417:Cattle; D003001:Cloning, Molecular; D017931:DNA Primers; D015342:DNA Probes; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D000072396:Endothelin-Converting Enzymes; D004730:Endothelium, Vascular / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D008666:Metalloendopeptidases; D008969:Molecular Sequence Data; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014357:Trypsin
D017931:DNA Primers; D015342:DNA Probes; D018076:DNA, Complementary; D011994:Recombinant Proteins; D014357:Trypsin; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; C000606923:ECE1 protein, human; D000072396:Endothelin-Converting Enzymes