Renal Angiotensin I-Converting Enzyme as a Mixture of Sialo- and Asialo-enzyme, and a Rapid Purification Method

Angiotensin I-converting enzyme [EC 3.4.15.1] was rapidly and highly purified from a particulate fraction of hog kidney cortex with 13% yield. The procedure, which was rapid, included fractionation on DEAE-cellulose and calcium phosphate gel, chromatographies on DEAE-Sephadex A-50 and hydroxylapatit...

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Bibliographic Details
Published in:Journal of biochemistry (Tokyo) 1976-09, Vol.80 (3), p.477-483
Main Authors: OSHIMA, Genichiro, NAGASAWA, Kinzo, KATO, Jyoji
Format: Article
Language:English
Subjects:
Animals
Chlorides - pharmacology
Chymotrypsin - pharmacology
Halogens - pharmacology
Hot Temperature
Hydrogen-Ion Concentration
Kidney - enzymology
Neuraminidase - metabolism
Peptidyl-Dipeptidase A - isolation & purification
Peptidyl-Dipeptidase A - metabolism
Sialic Acids
Swine
Trypsin - pharmacology
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Summary:Angiotensin I-converting enzyme [EC 3.4.15.1] was rapidly and highly purified from a particulate fraction of hog kidney cortex with 13% yield. The procedure, which was rapid, included fractionation on DEAE-cellulose and calcium phosphate gel, chromatographies on DEAE-Sephadex A-50 and hydroxylapatite columns, and gel filtration on a Sephadex G-200 column. The purified enzyme preparation gave two protein bands on standard disc gel electrophoresis, but showed a single protein component on the gel after treatment with neuraminidase [EC 3.2.1.18]. The data strongly suggest that the purified enzyme preparation was a mixture of sialo- and asialo-enzyme. Sialic acid residues apparently do not contribute to the catalytic activity of the enzyme. The enzyme was activated more by chloride ions than by other halide ions tested, using Bz-Gly-Gly-Gly as a substrate. The dissociation constant for chloride ions was determined to be 2.2 mM. Chloride did not protect the enzyme against heat or low pH. The enzyme was resistant to inactivation by trypsin [EC 3. 4. 21. 4] and chymotrypsin [EC 3.4.21.1].
ISSN:0021-924X
1756-2651
DOI:10.1093/oxfordjournals.jbchem.a131301