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Purification and characterization of pig lung carbonyl reductase.
Arch Biochem Biophys. 1992 Feb 01; 292(2):539-47.AB

Abstract

A pyrazole-sensitive carbonyl reductase from pig lung was purified to homogeneity by electrophoretic criteria. Chemical cross-linking study suggested that the native enzyme is a tetramer with a Mr of 103,000, consisting of apparent identical subunits of Mr 24,000. The enzyme reduced aliphatic and aromatic carbonyl compounds with NADPH as a preferable cofactor to NADH and catalyzed the oxidation of secondary alcohols and the aldehyde dismutation in the presence of NAD(P)+. Immunohistochemical study with the antibodies against the enzyme revealed that the enzyme was localized in the ciliated cells, nonciliated bronchiolar cells, Type II alveolar pneumocytes, and the epithelial cells of the ducts of the bronchial glands in the pig lung. In addition to the properties and distribution, the pig lung enzyme was immunochemically similar to the pulmonary enzymes in the guinea pig and mouse. However, the pig enzyme showed the following unusual features. (1) The enzyme exhibited an equatorial specificity in the reduction of 3-ketosteroids; the 4-pro-S hydrogen of NADPH was transferred to the carbonyl carbon atom of 5 alpha- and 5 beta-androstanes, and the respective reduced products were identified as 3 beta- and 3 alpha-hydroxysteroids. (2) Although the NADPH-linked reduction of carbonyl compounds apparently obeyed the Michaelis-Menten kinetics at pH 6.0, the double-reciprocal plots of the velocity vs concentrations of the carbonyl substrates were convex at pH higher than 6.5. The Hill coefficients and [S]0.5 values for the substrates decreased as the pH for reaction increased. The results suggest that the pig enzyme exhibits negative cooperativity with respect to the carbonyl substrates and that the hydrogen ion acts as an allosteric effector abolishing the negative interaction.

Authors+Show Affiliations

Department of Biochemistry, Gifu Pharmaceutical University, Japan.No affiliation info availableNo affiliation info availableNo 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

1731616

Citation

Oritani, H, et al. "Purification and Characterization of Pig Lung Carbonyl Reductase." Archives of Biochemistry and Biophysics, vol. 292, no. 2, 1992, pp. 539-47.
Oritani H, Deyashiki Y, Nakayama T, et al. Purification and characterization of pig lung carbonyl reductase. Arch Biochem Biophys. 1992;292(2):539-47.
Oritani, H., Deyashiki, Y., Nakayama, T., Hara, A., Sawada, H., Matsuura, K., Bunai, Y., & Ohya, I. (1992). Purification and characterization of pig lung carbonyl reductase. Archives of Biochemistry and Biophysics, 292(2), 539-47.
Oritani H, et al. Purification and Characterization of Pig Lung Carbonyl Reductase. Arch Biochem Biophys. 1992 Feb 1;292(2):539-47. PubMed PMID: 1731616.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Purification and characterization of pig lung carbonyl reductase. AU - Oritani,H, AU - Deyashiki,Y, AU - Nakayama,T, AU - Hara,A, AU - Sawada,H, AU - Matsuura,K, AU - Bunai,Y, AU - Ohya,I, PY - 1992/2/1/pubmed PY - 1992/2/1/medline PY - 1992/2/1/entrez SP - 539 EP - 47 JF - Archives of biochemistry and biophysics JO - Arch. Biochem. Biophys. VL - 292 IS - 2 N2 - A pyrazole-sensitive carbonyl reductase from pig lung was purified to homogeneity by electrophoretic criteria. Chemical cross-linking study suggested that the native enzyme is a tetramer with a Mr of 103,000, consisting of apparent identical subunits of Mr 24,000. The enzyme reduced aliphatic and aromatic carbonyl compounds with NADPH as a preferable cofactor to NADH and catalyzed the oxidation of secondary alcohols and the aldehyde dismutation in the presence of NAD(P)+. Immunohistochemical study with the antibodies against the enzyme revealed that the enzyme was localized in the ciliated cells, nonciliated bronchiolar cells, Type II alveolar pneumocytes, and the epithelial cells of the ducts of the bronchial glands in the pig lung. In addition to the properties and distribution, the pig lung enzyme was immunochemically similar to the pulmonary enzymes in the guinea pig and mouse. However, the pig enzyme showed the following unusual features. (1) The enzyme exhibited an equatorial specificity in the reduction of 3-ketosteroids; the 4-pro-S hydrogen of NADPH was transferred to the carbonyl carbon atom of 5 alpha- and 5 beta-androstanes, and the respective reduced products were identified as 3 beta- and 3 alpha-hydroxysteroids. (2) Although the NADPH-linked reduction of carbonyl compounds apparently obeyed the Michaelis-Menten kinetics at pH 6.0, the double-reciprocal plots of the velocity vs concentrations of the carbonyl substrates were convex at pH higher than 6.5. The Hill coefficients and [S]0.5 values for the substrates decreased as the pH for reaction increased. The results suggest that the pig enzyme exhibits negative cooperativity with respect to the carbonyl substrates and that the hydrogen ion acts as an allosteric effector abolishing the negative interaction. SN - 0003-9861 UR - https://www.unboundmedicine.com/medline/citation/1731616/Purification_and_characterization_of_pig_lung_carbonyl_reductase_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0003-9861(92)90028-U DB - PRIME DP - Unbound Medicine ER -