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Carbonyl reductase of dog liver: purification, properties, and kinetic mechanism.
Arch Biochem Biophys. 1986 Jan; 244(1):238-47.AB

Abstract

A carbonyl reductase has been extracted into 0.5 M KCl from dog liver and purified to apparent homogeneity by a three-step procedure consisting of chromatography on CM-Sephadex, Matrex green A, and Sephadex G-100 in high-ionic-strength buffers. The enzyme is a dimer composed of two identical subunits of molecular weight 27,000. The pH optimum is 5.5 and the isoelectric point of the enzyme is 9.3. The enzyme reduces aromatic ketones and aldehydes; the aromatic ketones with adjacent medium alkyl chains are the best substrates. Quinones, ketosteroids, prostaglandins, and aliphatic carbonyl compounds are poor or inactive substrates for the enzyme. As a cofactor the enzyme utilizes NADPH, the pro-S hydrogen atom of which is transferred to the substrate. Two moles of NADPH bind to one mole of the enzyme molecule, causing a blue shift and enhancement of the cofactor fluorescence. The reductase reaction is reversible and the equilibrium constant determined at pH 7.0 is 12.8. Steady-state kinetic measurements in both directions suggest that the reaction proceeds through a di-iso ordered bi-bi mechanism.

Authors

No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

3511844

Citation

Hara, A, et al. "Carbonyl Reductase of Dog Liver: Purification, Properties, and Kinetic Mechanism." Archives of Biochemistry and Biophysics, vol. 244, no. 1, 1986, pp. 238-47.
Hara A, Nakayama T, Deyashiki Y, et al. Carbonyl reductase of dog liver: purification, properties, and kinetic mechanism. Arch Biochem Biophys. 1986;244(1):238-47.
Hara, A., Nakayama, T., Deyashiki, Y., Kariya, K., & Sawada, H. (1986). Carbonyl reductase of dog liver: purification, properties, and kinetic mechanism. Archives of Biochemistry and Biophysics, 244(1), 238-47.
Hara A, et al. Carbonyl Reductase of Dog Liver: Purification, Properties, and Kinetic Mechanism. Arch Biochem Biophys. 1986;244(1):238-47. PubMed PMID: 3511844.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Carbonyl reductase of dog liver: purification, properties, and kinetic mechanism. AU - Hara,A, AU - Nakayama,T, AU - Deyashiki,Y, AU - Kariya,K, AU - Sawada,H, PY - 1986/1/1/pubmed PY - 1986/1/1/medline PY - 1986/1/1/entrez SP - 238 EP - 47 JF - Archives of biochemistry and biophysics JO - Arch. Biochem. Biophys. VL - 244 IS - 1 N2 - A carbonyl reductase has been extracted into 0.5 M KCl from dog liver and purified to apparent homogeneity by a three-step procedure consisting of chromatography on CM-Sephadex, Matrex green A, and Sephadex G-100 in high-ionic-strength buffers. The enzyme is a dimer composed of two identical subunits of molecular weight 27,000. The pH optimum is 5.5 and the isoelectric point of the enzyme is 9.3. The enzyme reduces aromatic ketones and aldehydes; the aromatic ketones with adjacent medium alkyl chains are the best substrates. Quinones, ketosteroids, prostaglandins, and aliphatic carbonyl compounds are poor or inactive substrates for the enzyme. As a cofactor the enzyme utilizes NADPH, the pro-S hydrogen atom of which is transferred to the substrate. Two moles of NADPH bind to one mole of the enzyme molecule, causing a blue shift and enhancement of the cofactor fluorescence. The reductase reaction is reversible and the equilibrium constant determined at pH 7.0 is 12.8. Steady-state kinetic measurements in both directions suggest that the reaction proceeds through a di-iso ordered bi-bi mechanism. SN - 0003-9861 UR - https://www.unboundmedicine.com/medline/citation/3511844/Carbonyl_reductase_of_dog_liver:_purification_properties_and_kinetic_mechanism_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0003-9861(86)90113-X DB - PRIME DP - Unbound Medicine ER -