The endopeptidase activity and the activation by Cl- of angiotensin-converting enzyme is evolutionarily conserved: purification and properties of an an angiotensin-converting enzyme from the housefly, Musca domestica.
A soluble 67 kDa angiotensin-converting enzyme (ACE) has been purified by lisinopril-Sepharose affinity column chromatography from adult houseflies, Musca domestica. The dipeptidyl carboxypeptidase activity towards benzoyl-Gly-His-Leu was inhibited by captopril (IC50 50 nM) and fosinoprilat (IC50 251 nM), two inhibitors of mammalian ACE, and was activated by Cl- (optimal Cl- concentration 600 mM). Musca ACE removed C-terminal dipeptides from angiotensin I, bradykinin [Leu5]enkephalin and [Met5]enkephalin and also functioned as an endopeptidase by hydrolysing dipeptideamides from [Leu5]enkephalinamide and [Met5]enkephalinamide, and a dipeptideamide and a tripeptideamide from substance P. Musca ACE was also able to cleave a tripeptide from both the N-terminus and C-terminus of luteinizing hormone-releasing hormone, with C-terminal hydrolysis predominating. Maximal N-terminal tripeptidase activity occurred at 150 mM NaCl, whereas the C-terminal tripeptidase activity continued to rise with increasing concentration of Cl- (0-0.5 M). Musca ACE displays properties of both the N- and C-domains of human ACE, indicating a high degree of conservation during evolution of the substrate specificity of ACE and its response to Cl-.
Lamango
N S
NS
Department of Biology, University of Leeds, U.K.
Sajid
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M
Isaac
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RE
D000595:Amino Acid Sequence; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000494:pharmacology; D000818:Animals; D005075:Biological Evolution; D002846:Chromatography, Affinity; D017124:Conserved Sequence; D010450:Endopeptidases / Q000378:metabolism*; D004789:Enzyme Activation; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism; D006793:Houseflies / Q000201:enzymology*; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D007703:Peptidyl-Dipeptidase A / Q000302:isolation & purification* / Q000378:metabolism*; D012965:Sodium Chloride / Q000494:pharmacology*
D000806:Angiotensin-Converting Enzyme Inhibitors; D010455:Peptides; D007987:Gonadotropin-Releasing Hormone; D012965:Sodium Chloride; D010450:Endopeptidases; D007703:Peptidyl-Dipeptidase A