TY - JOUR
T1 - [71] Steroid-Transforming Enzymes
AU - Benson, Ann M.
AU - Suruda, Anthony J.
AU - Barrack, Evelyn R.
AU - Talalay, Paul
N1 - Funding Information:
Acknowledgment These studies were supported by Research Grant AM07422 from the U.S. Public Health Service and by a Henry Strong Denison Fellowship of The Johns Hopkins University School of Medicine (A.J.S.).
Copyright:
Copyright 2018 Elsevier B.V., All rights reserved.
PY - 1974/1/1
Y1 - 1974/1/1
N2 - Affinity chromatography on adsorbents containing steroidal substrate analogs covalently linked to agarose promises to be an extremely valuable technique both for the purification of steroid-transforming enzymes, and for studies on their mechanism. When Pseudomonas testosteroni is grown on media containing certain steroids, crude extracts of lyophilized or sonically disrupted cells are a rich source of various induced steroid transforming enzymes. The isolation from these mixtures of discrete catalytic proteins, in purified form and uncontaminated by each other, has been a challenging task. The availability of such purified enzymes would greatly augment their suitability for the microanalysis and determination of steric configuration of steroids. Affinity techniques have indicated that the fl-enzymes do not bind strongly to steroids in the absence of NAD+, thus suggesting that these oxidations may involve an obligatory ordered addition in which the binding of NAD+ precedes that of the steroidal substrate.
AB - Affinity chromatography on adsorbents containing steroidal substrate analogs covalently linked to agarose promises to be an extremely valuable technique both for the purification of steroid-transforming enzymes, and for studies on their mechanism. When Pseudomonas testosteroni is grown on media containing certain steroids, crude extracts of lyophilized or sonically disrupted cells are a rich source of various induced steroid transforming enzymes. The isolation from these mixtures of discrete catalytic proteins, in purified form and uncontaminated by each other, has been a challenging task. The availability of such purified enzymes would greatly augment their suitability for the microanalysis and determination of steric configuration of steroids. Affinity techniques have indicated that the fl-enzymes do not bind strongly to steroids in the absence of NAD+, thus suggesting that these oxidations may involve an obligatory ordered addition in which the binding of NAD+ precedes that of the steroidal substrate.
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U2 - 10.1016/S0076-6879(74)34074-8
DO - 10.1016/S0076-6879(74)34074-8
M3 - Article
C2 - 4375238
AN - SCOPUS:0016142534
VL - 34
SP - 557
EP - 566
JO - Methods in Enzymology
JF - Methods in Enzymology
SN - 0076-6879
IS - C
ER -