Advertisement
JBC

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


     

Originally published In Press as doi:10.1074/jbc.M213168200 on January 30, 2003

J. Biol. Chem., Vol. 278, Issue 15, 13196-13206, April 11, 2003
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
278/15/13196    most recent
M213168200v1
Right arrow Submit a Letter to Editor
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
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 arrowRequest Permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Darbandi-Tonkabon, R.
Right arrow Articles by Evers, A. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Darbandi-Tonkabon, R.
Right arrow Articles by Evers, A. S.
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?

Photoaffinity Labeling with a Neuroactive Steroid Analogue
6-AZI-PREGNANOLONE LABELS VOLTAGE-DEPENDENT ANION CHANNEL-1 IN RAT BRAIN*

Ramin Darbandi-TonkabonDagger , William R. HastingsDagger , Chun-Min Zeng§, Gustav AkkDagger , Brad D. ManionDagger , John R. BracamontesDagger , Joseph H. SteinbachDagger , Steven J. Mennerick, Douglas F. Covey§, and Alex S. EversDagger §||

From the Departments of Dagger  Anesthesiology, § Molecular Biology and Pharmacology, and  Psychiatry, Washington University School of Medicine, St. Louis, Missouri 63110

Neuroactive steroids modulate the function of gamma -aminobutyric acid, type A (GABAA) receptors in the central nervous system by an unknown mechanism. In this study we have used a novel neuroactive steroid analogue, 3alpha ,5beta -6-azi-3-hydroxypregnan-20-one (6-AziP), as a photoaffinity labeling reagent to identify neuroactive steroid binding sites in rat brain. 6-AziP is an effective modulator of GABAA receptors as evidenced by its ability to inhibit binding of [35S]t-butylbicyclophosphorothionate to rat brain membranes and to potentiate GABA-elicited currents in Xenopus oocytes and human endothelial kidney 293 cells expressing GABAA receptor subunits (alpha 1beta 2gamma 2). [3H]6-AziP produced time- and concentration-dependent photolabeling of protein bands of ~35 and 60 kDa in rat brain membranes. The 35-kDa band was half-maximally labeled at a [3H]6-AziP concentration of 1.9 µM, whereas the 60-kDa band was labeled at higher concentrations. The photolabeled 35-kDa protein was isolated from rat brain by two-dimensional PAGE and identified as voltage-dependent anion channel-1 (VDAC-1) by both matrix-assisted laser desorption ionization time-of-flight and ESI-tandem mass spectrometry. Monoclonal antibody directed against the N terminus of VDAC-1 immunoprecipitated labeled 35-kDa protein from a lysate of rat brain membranes, confirming that VDAC-1 is the species labeled by [3H]6-AziP. The beta 2 and beta 3 subunits of the GABAA receptor were co-immunoprecipitated by the VDAC-1 antibody suggesting a physical association between VDAC-1 and GABAA receptors in rat brain membranes. These data suggest that neuroactive steroid effects on the GABAA receptor may be mediated by binding to an accessory protein, VDAC-1.

* This work was supported in part by National Institutes of Health Grants PO1-GM47969 (to J. H. S., D. F. C., and A. S. E.) and AA12952 (to S. J. M.) and by a grant from the Klingenstein Foundation (to S. J. M.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

|| To whom correspondence should be addressed: Dept. of Anesthesiology, Washington University School of Medicine, 660 S. Euclid Ave., Campus Box 8054, St. Louis, MO 63110. Tel.: 314-454-8702; Fax: 314-454-5572; E-mail: eversa@notes.wustl.edu.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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
 J. Biol. Chem.Home page
J. R. Bracamontes and J. H. Steinbach
Multiple Modes for Conferring Surface Expression of Homomeric {beta}1 GABAA Receptors
J. Biol. Chem., September 19, 2008; 283(38): 26128 - 26136.
[Abstract] [Full Text] [PDF]

Home page
 J. Pharmacol. Exp. Ther.Home page
T. Bordet, B. Buisson, M. Michaud, C. Drouot, P. Galea, P. Delaage, N. P. Akentieva, A. S. Evers, D. F. Covey, M. A. Ostuni, et al.
Identification and Characterization of Cholest-4-en-3-one, Oxime (TRO19622), a Novel Drug Candidate for Amyotrophic Lateral Sclerosis
J. Pharmacol. Exp. Ther., August 1, 2007; 322(2): 709 - 720.
[Abstract] [Full Text] [PDF]

Home page
 Endocr. Rev.Home page
M. Schumacher, R. Guennoun, A. Ghoumari, C. Massaad, F. Robert, M. El-Etr, Y. Akwa, K. Rajkowski, and E.-E. Baulieu
Novel Perspectives for Progesterone in Hormone Replacement Therapy, with Special Reference to the Nervous System
Endocr. Rev., June 1, 2007; 28(4): 387 - 439.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
D. P. Bright, S. D. Adham, L. C. J. M. Lemaire, R. Benavides, M. Gruss, G. W. Taylor, E. H. Smith, and N. P. Franks
Identification of Anesthetic Binding Sites on Human Serum Albumin Using a Novel Etomidate Photolabel
J. Biol. Chem., April 20, 2007; 282(16): 12038 - 12047.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. ProteomicsHome page
I. Roman, J. Figys, G. Steurs, and M. Zizi
Hunting Interactomes of a Membrane Protein: Obtaining the Largest Set of Voltage-Dependent Anion Channel-Interacting Protein Epitopes
Mol. Cell. Proteomics, September 1, 2006; 5(9): 1667 - 1680.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. Xi, R. Liu, G. R. Asbury, M. F. Eckenhoff, and R. G. Eckenhoff
Inhalational Anesthetic-binding Proteins in Rat Neuronal Membranes
J. Biol. Chem., May 7, 2004; 279(19): 19628 - 19633.
[Abstract] [Full Text] [PDF]

Home page
 J. Pharmacol. Exp. Ther.Home page
R. Darbandi-Tonkabon, B. D. Manion, W. R. Hastings, W. J. Craigen, G. Akk, J. R. Bracamontes, Y. He, T. V. Sheiko, J. H. Steinbach, S. J. Mennerick, et al.
Neuroactive Steroid Interactions with Voltage-Dependent Anion Channels: Lack of Relationship to GABAA Receptor Modulation and Anesthesia
J. Pharmacol. Exp. Ther., February 1, 2004; 308(2): 502 - 511.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2003 by the American Society for Biochemistry and Molecular Biology.
Advertisement
spacer
Advertisement
Advertisement