Biomaterial Database

"Affinity" chromatography of steroid-transforming enzymes with a non-steroidal ligand. 1979

A G Renwick, and S M Chambers, and P Willcox

The chromatographic behaviour of an avian oestradiol-17 beta dehydrogenase, the 3(17) beta-hydroxy steroid dehydrogenase from Pseudomonas testosteroni and cortisone reductase from Streptomyces dehydrogenans was studied on columns of p-(phenoxypropoxy)aniline attached to CNBr-activated Sepharose. The ligand was effective in adsorbing the oestradiol dehydrogenase from a partially purified extract of chicken liver, and the cortisone reductase was perferentially retained when mixtures of the three dehydrogenases were applied to columns in 10mM-buffer. Under these conditions the 3(17)beta-hydroxy steroid dehydrogenase was eluted in the front, but was adsorbed in the presence of 3 M-KCl. beta-N-Acetylglucosaminidase present in the liver preparation was not retained by the ligand, whereas lactate dehydrogenase from rabbit muscle was adsorbed in a manner similar to the retention pattern found on affinity chromatography with 2',5'-ADP--Sepharose. The mean overall purification of the oestradiol dehydrogenase was 13-fold, with a mean recovery of 53%. p-(Phenoxypropoxy)aniline offers promise for the purification of steroid-transforming enzymes where elution with substrate or cofactor is not wanted. It is also suggested that the ligand may be of service in the purification of receptors of hormonal steroids.

UI	MeSH Term	Description	Entries
D007770	L-Lactate Dehydrogenase	A tetrameric enzyme that, along with the coenzyme NAD+, catalyzes the interconversion of LACTATE and PYRUVATE. In vertebrates, genes for three different subunits (LDH-A, LDH-B and LDH-C) exist.	Lactate Dehydrogenase,Dehydrogenase, L-Lactate,Dehydrogenase, Lactate,L Lactate Dehydrogenase
D008024	Ligands	A molecule that binds to another molecule, used especially to refer to a small molecule that binds specifically to a larger molecule, e.g., an antigen binding to an antibody, a hormone or neurotransmitter binding to a receptor, or a substrate or allosteric effector binding to an enzyme. Ligands are also molecules that donate or accept a pair of electrons to form a coordinate covalent bond with the central metal atom of a coordination complex. (From Dorland, 27th ed)	Ligand
D002846	Chromatography, Affinity	A chromatographic technique that utilizes the ability of biological molecules, often ANTIBODIES, to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)	Chromatography, Bioaffinity,Immunochromatography,Affinity Chromatography,Bioaffinity Chromatography
D003349	Cortisone Reductase	An enzyme that catalyzes the interconversion of a ketone and hydroxy group at C-20 of cortisone and other 17,20,21-trihydroxy steroids. EC 1.1.1.53.	20-beta-Hydroxysteroid Dehydrogenase,3 alpha,20 beta Hydroxysteroid Dehydrogenase,20 beta Hydroxysteroid Dehydrogenase,Dehydrogenase, 20-beta-Hydroxysteroid,Reductase, Cortisone
D004960	Estradiol Dehydrogenases	Enzymes that catalyze the oxidation of estradiol at the 17-hydroxyl group in the presence of NAD+ or NADP+ to yield estrone and NADH or NADPH. The 17-hydroxyl group can be in the alpha- or beta-configuration. EC 1.1.1.62	17 beta-Estradiol Dehydrogenase,Estradiol 17 alpha-Dehydrogenase,Estradiol 17 beta-Dehydrogenase,Estradiol Dehydrogenase,17 alpha-Dehydrogenase, Estradiol,17 beta Estradiol Dehydrogenase,17 beta-Dehydrogenase, Estradiol,Dehydrogenase, 17 beta-Estradiol,Dehydrogenase, Estradiol,Dehydrogenases, Estradiol,Estradiol 17 alpha Dehydrogenase,Estradiol 17 beta Dehydrogenase
D000118	Acetylglucosaminidase	A beta-N-Acetylhexosaminidase that catalyzes the hydrolysis of terminal, non-reducing 2-acetamido-2-deoxy-beta-glucose residues in chitobiose and higher analogs as well as in glycoproteins. Has been used widely in structural studies on bacterial cell walls and in the study of diseases such as MUCOLIPIDOSIS and various inflammatory disorders of muscle and connective tissue.	N-Acetyl-beta-D-glucosaminidase,Chitobiase,N,N-Diacetylchitobiase,N-Ac-beta-Glucosaminidase,NAGase,beta-D-Acetamido-2-Deoxyglucosidase,beta-D-N-acetylglucosaminidase,beta-N-Acetylglucosaminidase,N Ac beta Glucosaminidase,N Acetyl beta D glucosaminidase,N,N Diacetylchitobiase,beta D Acetamido 2 Deoxyglucosidase,beta D N acetylglucosaminidase,beta N Acetylglucosaminidase
D000814	Aniline Compounds	Compounds that include the aminobenzene structure.	Phenylamine,Phenylamines,Anilines,Compounds, Aniline
D015067	17-Hydroxysteroid Dehydrogenases	A class of enzymes that catalyzes the oxidation of 17-hydroxysteroids to 17-ketosteroids. EC 1.1.-.	17 beta-Hydroxysteroid Dehydrogenases,Testosterone Dehydrogenases,17-Ketosteroid Reductases,17 Hydroxysteroid Dehydrogenases,17 Ketosteroid Reductases,17 beta Hydroxysteroid Dehydrogenases,Dehydrogenases, 17 beta-Hydroxysteroid,Dehydrogenases, 17-Hydroxysteroid,Dehydrogenases, Testosterone,Reductases, 17-Ketosteroid
D015089	20-Hydroxysteroid Dehydrogenases	A group of enzymes that catalyze the reversible reduction-oxidation reaction of 20-hydroxysteroids, such as from a 20-ketosteroid to a 20-alpha-hydroxysteroid (EC 1.1.1.149) or to a 20-beta-hydroxysteroid (EC 1.1.1.53).	20 Hydroxysteroid Dehydrogenases,Dehydrogenases, 20 Hydroxysteroid,Dehydrogenases, 20-Hydroxysteroid,Hydroxysteroid Dehydrogenases, 20