Purification of soluble and membrane forms of somatic angiotensin-converting enzyme by cascade affinity chromatography.
Soluble and membrane forms of angiotensin-converting enzyme were purified by cascade affinity chromatography. The enzyme forms were completely separated from each other using their different affinity to the hydrophobic matrix phenyl-silochrome. The enzymes was further purified on affinity sorbent prepared by immobilization of the enzyme inhibitor N-[1(S)-carboxy-5-aminopentyl]glycylphenylalanine on agarose. The procedure yielded electrophoretically homogeneous soluble and membrane forms of angiotensin-converting enzyme containing only active molecules as demonstrated by titration with the reversible inhibitor lisinopril. According to phase separation in the presence of Triton X-114, the membrane enzyme is more hydrophobic than the soluble form. The catalytic characteristics of the enzyme forms differed from each other in the system Aerosol OT-water-octane (reversed micelles) which is model for the membrane environment of the enzymes in vivo.
Kost
O A
OA
School of Chemistry, Lomonosov Moscow State University, Russia. kost@enzyme.chem.msu.su
Grinshtein
S V
SV
Nikolskaya
I I
II
Shevchenko
A A
AA
Binevski
P V
PV
D000806:Angiotensin-Converting Enzyme Inhibitors / Q000494:pharmacology; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D003600:Cytosol / Q000201:enzymology; D004151:Dipeptides; D004800:Enzymes, Immobilized; D007527:Isoenzymes / Q000302:isolation & purification; D017706:Lisinopril / Q000494:pharmacology; D008168:Lung / Q000201:enzymology; D008565:Membrane Proteins / Q000302:isolation & purification; D008823:Micelles; D017830:Octoxynol; D007703:Peptidyl-Dipeptidase A / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D011092:Polyethylene Glycols; D012995:Solubility
D000806:Angiotensin-Converting Enzyme Inhibitors; D004151:Dipeptides; D004800:Enzymes, Immobilized; D007527:Isoenzymes; D008565:Membrane Proteins; D008823:Micelles; C108065:N-(1-carboxy-5-aminopentyl)glycylphenylalanine; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40; D017706:Lisinopril; D007703:Peptidyl-Dipeptidase A