Open Access
Chemical Modification of Cysteinyl, Lysyl and Histidyl Residues of Mouse Liver 17β-Hydroxysteroid Dehydrogenase.
Nakayama,Toshihiro,Tanabe,Hiroyuki,Deyashiki,Yoshihiro,Shinoda,Michio,Hara,Akira,Sawada,Hideo +11 more
- Vol. 42, pp 66
Reads0
Chats0
TLDR
The results suggest the presence of essential cysteine and lysine residues at or near the coenzyme-binding site and that of essential histidine residue(s) in the catalytic region of the active site of mouse liver 17 beta-hydroxysteroid dehydrogenase.Abstract:
Monomeric 17 beta-hydroxysteroid dehydrogenase from mouse liver was rapidly inactivated by 5,5'-dithiobis(2-nitrobenzoic acid) and 2,4,6-trinitrobenzene-1-sulfonate, and the absorption spectra of the inactivated enzymes indicated that cysteine and lysine residues were modified. The kinetics of inactivation and spectrophotometric quantification of the modified residues suggested that complete inactivation was caused by modification of two cysteine residues or one lysine residue per active site. The inactivation by the two reagents was protected by NADP+ and some coenzyme analogs, but not by a steroid substrate, testosterone. Moreover, chemical modification by diethyl pyrocarbonate also produced inactivation of the enzyme, and showed a difference spectrum with a peak at 242 nm characteristic of N-carbethoxyhistidine residues, which decreased with the addition of hydroxylamine. The inactivation by this reagent, following pseudo-first-order kinetics, was protected partially by either NADP+ or testosterone and completely in the presence of both the coenzyme and substrate. The results suggest the presence of essential cysteine and lysine residues at or near the coenzyme-binding site and that of essential histidine residue(s) in the catalytic region of the active site of mouse liver 17 beta-hydroxysteroid dehydrogenase.read more
Citations
More filters
Mouse Liver Dihydrodiol Dehydrogenases. : Identity of the Predominant and a Minor Form with 17β-Hydroxysteroid Dehydrogenase and Aldehyde Reductase
Sawada,Hideo,Hara,Akira,Nakayama,Toshihiro,Nakagawa,Makoto,Inoue,Yoshio,Hasebe,Kazuya,Zhang,Yuan-Pei +13 more
TL;DR: The results indicate that the major form of dihydrodiol dehydrogenase is identical to 17 beta-hydroxysteroid dehydrogen enzyme and the minor enzyme form to aldehyde reductase, which reduced various aldehydes well and was specifically inhibited by barbiturates and sorbinil.
Book
Enzymology and Molecular Biology of Carbonyl Metabolism 6
TL;DR: Three-Dimensional Structures of Human Alcohol Dehydrogenase Isoenzymes Reveal the Molecular basis for Their Functional Diversity and three-D dimensional structures of Human Corneal and Lens Aldehyde dehydrogenases reveal the molecular basis for their Functional Diversity.
Journal ArticleDOI
Characterization of fungal 17β-hydroxysteroid dehydrogenases
TL;DR: There are most probably different enzymes responsible for 17β-HSD activity in filamentous fungi, according to the nature of these enzymes investigated.
Journal ArticleDOI
Redox regulation of human estrogen sulfotransferase (hSULT1E1).
TL;DR: The redox regulation of hSULT1E1 may interrupt the regulation and function of estrogens under various physiological and pathological conditions, according to this first report on the redoxregulation of human SULTs.
Kinetic and Stereochemical Studies on Reaction Mechanism of Mouse Liver 17β-Hydroxysteroid Dehydrogenases.
TL;DR: The kinetic mechanism of two major monomeric 17β-hydroxysteroid dehydrogenases from mouse liver cytosol was studied at pH 7 in both directions with NADP(H) and three steroid substrates: testosterone, 5 beta-androstane-3 alpha, 17 17 beta-diol, and estradiol-17 beta.
References
More filters
Book ChapterDOI
[37] Reaction of protein sulfhydryl groups with Ellman's reagent.
TL;DR: This chapter discusses a reaction of protein sulfhydryl groups with Ellman's reagent 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), found to be a sensitive tool for the assay of thiol groups in tissues, body fluids, and proteins.
Book ChapterDOI
Modification of histidyl residues in proteins by diethylpyrocarbonate.
TL;DR: Although diethylpyrocarbonate does not always react specifically with histidyl residues in proteins, it is more selective than other acylating agents and can give useful information about the role of histidol residues in many proteins.
Journal ArticleDOI
Inactivation of Myosin by 2,4-Dinitrophenol and Protection by Adenosine Triphosphate and Other Phosphate Compounds
TL;DR: It has become apparent that adenosine triphosphate binds to the active site of myosin in a way which is not optimal for hydrolytic cleavage of the terminal phosphate bond, and a wide variety of reagents, including 2,4-dinitrophenol, can improve the catalysis of this reaction by altering the interaction between enzyme and substrate.
Journal ArticleDOI
On the preparation and properties of 2, 4, 6-trinitrophenyl-amino acids and-peptides
Tsuneo Okuyama,Kazuo Satake +1 more
Journal ArticleDOI
Cloning and sequencing of the cDNA for rat liver 3 alpha-hydroxysteroid/dihydrodiol dehydrogenase.
TL;DR: The sequence data presented suggests that 3 alpha-HSD, prostaglandin F synthase, and aldehyde/aldose reductases are members of a common gene family.
Related Papers (5)
Effect of trinitrophenylation of specific lysyl residues on the catalytic, regulatory, and molecular properties of bovine liver glutamate dehydrogenase.
Goldin Br,Frieden C +1 more
Chemical modification of amino acid residues essential for bovine heart 2-oxoglutarate dehydrogenase activity.
Reversible, covalent modification of the essential cysteine residues of pig heart lactate dehydrogenase
J. J. Holbrook,Renate Jeckel +1 more