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Purification of soluble and membrane forms of somatic angiotensin-converting enzyme by cascade affinity
O A Kost1, S V Grinshtein, I I Nikolskaya
1School of Chemistry, Lomonosov Moscow State University, Russia. kost@enzyme.chem.msu.su
Abstract:
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.