JBC Oz Biosciences

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Furster, C.
Right arrow Articles by Toll, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Furster, C.
Right arrow Articles by Toll, A.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Volume 271, Number 34, Issue of August 23, 1996 pp. 20903-20907
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

Purification of a 3beta -Hydroxy-Delta 5-C27-steroid Dehydrogenase from Pig Liver Microsomes Active in Major and Alternative Pathways of Bile Acid Biosynthesis

(Received for publication, May 13, 1996)

Catrin Furster Dagger , Jie Zhang and Anders Toll Dagger

From the Dagger  Division of Biochemistry, Department of Pharmaceutical Biosciences, University of Uppsala, Uppsala and  Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden

A 3beta -hydroxy-Delta 5-C27-steroid dehydrogenase active in bile acid biosynthesis was purified from pig liver microsomes by solubilization with sodium cholate and by chromatography on DEAE-Sepharose, aminohexyl-Sepharose, and blue Sepharose. The last step in the purification procedure was preparative isoelectric focusing in a Rotofor cell. The final enzyme preparation showed only one protein band upon SDS-polyacrylamide gel electrophoresis. The isoelectric point was estimated to about 7.0 and the apparent Mr was 36,000.

The purified enzyme catalyzed the conversion of 7alpha -hydroxycholesterol, 7alpha ,25-dihydroxycholesterol, 7alpha ,27-dihydroxycholesterol, and 3beta ,7alpha -dihydroxy-5-cholestenoic acid into the corresponding 3-oxo-Delta 4 compounds. The enzyme was inactive with C19 and C21 steroids as substrates. The enzyme was also inactive with C27 steroids having the 7-hydroxy group in beta - instead of alpha -position. The Km was found to be 0.30 and 0.32 µM with 7alpha -hydroxycholesterol and 7alpha ,27-dihydroxycholesterol as substrates, respectively. NAD+ was the preferred cofactor. A monoclonal antibody raised against the 3beta -hydroxy-Delta 5-C27-steroid dehydrogenase was prepared. After coupling to Sepharose, the antibody was able to bind the dehydrogenase and to decrease the conversion of 7alpha -hydroxycholesterol into 7alpha -hydroxy-4-cholest-3-one by more than 90%. The N-terminal amino acid sequence was determined and found to be similar but not identical with those of known 3beta -hydroxy-Delta 5-steroid dehydrogenases active in steroid hormone biosynthesis. Thus, the purified enzyme active toward C27 steroids in bile acid biosynthesis appears to represent a novel type of 3beta -hydroxy-Delta 5-steroid dehydrogenase.


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 Endocr. Rev.Home page
J. Simard, M.-L. Ricketts, S. Gingras, P. Soucy, F. A. Feltus, and M. H. Melner
Molecular Biology of the 3{beta}-Hydroxysteroid Dehydrogenase/{Delta}5-{Delta}4 Isomerase Gene Family
Endocr. Rev., June 1, 2005; 26(4): 525 - 582.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
J. B. Cheng, E. Jacquemin, M. Gerhardt, H. Nazer, D. Cresteil, J. E. Heubi, K. D. R. Setchell, and D. W. Russell
Molecular Genetics of 3{beta}-Hydroxy-{Delta}5-C27-Steroid Oxidoreductase Deficiency in 16 Patients with Loss of Bile Acid Synthesis and Liver Disease
J. Clin. Endocrinol. Metab., April 1, 2003; 88(4): 1833 - 1841.
[Abstract] [Full Text] [PDF]

Home page
 J. Lipid Res.Home page
M. Norlin
Expression of key enzymes in bile acid biosynthesis during development: CYP7B1-mediated activities show tissue-specific differences
J. Lipid Res., May 1, 2002; 43(5): 721 - 731.
[Abstract] [Full Text] [PDF]

Home page
 J. Lipid Res.Home page
G. Alvelius, O. Hjalmarson, W. J. Griffiths, I. Bjorkhem, and J. Sjovall
Identification of unusual 7-oxygenated bile acid sulfates in a patient with Niemann-Pick disease, type C
J. Lipid Res., October 1, 2001; 42(10): 1571 - 1577.
[Abstract] [Full Text] [PDF]

Home page
 Drug Metab. Dispos.Home page
T. Matsunaga, N. Kishi, S. Higuchi, K. Watanabe, T. Ohshima, and I. Yamamoto
CYP3A4 Is a Major Isoform Responsible for Oxidation of 7-Hydroxy-Delta 8-tetrahydrocannabinol to 7-Oxo-Delta 8-tetrahydrocannabinol in Human Liver Microsomes
Drug Metab. Dispos., November 1, 2000; 28(11): 1291 - 1296.
[Abstract] [Full Text]

Home page
 J. Lipid Res.Home page
A. Babiker, O. Andersson, D. Lindblom, J. van der Linden, B. Wiklund, D. Lütjohann, U. Diczfalusy, and I. Björkhem
Elimination of cholesterol as cholestenoic acid in human lung by sterol 27-hydroxylase: evidence that most of this steroid in the circulation is of pulmonary origin
J. Lipid Res., August 1, 1999; 40(8): 1417 - 1425.
[Abstract] [Full Text]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 1996 by the American Society for Biochemistry and Molecular Biology.