Purification and characterization of a dipeptidyl carboxypeptidase from the polychaete Neanthes virens resembling angiotensin I converting enzyme.
Dipeptidyl carboxypeptidase (DCP) is well known as a mammalian angiotensin I converting enzyme (ACE) which plays an important role in blood pressure homeostasis. DCP was purified from the whole body of a polychaete, Neanthes virens. The purified enzyme was homogeneous by SDS-PAGE, with a molecular mass of 71 kDa by SDS-PAGE and 69 kDa by gel filtration, indicating that it is monomeric. The isoelectric point was 4.5 and optimum pH for the activity was 8.0. It showed a specific activity of 466.8 U/mg, which is the highest of known DCPs. The enzyme hydrolyzed angiotensin I to angiotensin II and sequentially released Phe-Arg and Ser-Pro from the C-terminus bradykinin, but does not cleave imido-bonds. Activity was completely inhibited by 1 mM EDTA and 5 mM o-phenanthroline, but it was not affected by serine and aspartic protease inhibitors. The original activity of EDTA-inactivated DCP was restored by addition of cobalt, manganese or low concentrations of zinc. The Km and Vmax values of the enzyme for Bz-Gly-His-Leu were 0.56 mM and 600 mumol/min per mg, respectively. The Ki values for specific mammalian ACE inhibitors, such as captopril and lisinopril, were 1.38 and 2.07 nM, respectively. In conclusion, we have shown the existence of a DCP from the polychaete, N. virens, with similar properties to those of mammalian ACE.
Kawamura
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T
Industrial Technology Center of Nagasaki, Japan.
Oda
T
T
Muramatsu
T
T
D000818:Animals; D002384:Catalysis; D002413:Cations, Divalent; D010450:Endopeptidases / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D007526:Isoelectric Point; D008670:Metals; D008970:Molecular Weight; D007703:Peptidyl-Dipeptidase A / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism*; D011077:Polychaeta / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature
D002413:Cations, Divalent; D007202:Indicators and Reagents; D008670:Metals; D010450:Endopeptidases; C020071:dipeptidyl carboxypeptidase; D007703:Peptidyl-Dipeptidase A