JBC PeproTech; Our Business is Cytokines!

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


     

Originally published In Press as doi:10.1074/jbc.M211292200 on February 3, 2003

J. Biol. Chem., Vol. 278, Issue 15, 12672-12679, April 11, 2003
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow An addition or correction has been published
Right arrow All Versions of this Article:
278/15/12672    most recent
M211292200v1
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 Gille, A.
Right arrow Articles by Seifert, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Gille, A.
Right arrow Articles by Seifert, R.
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?

2'(3')-O-(N-Methylanthraniloyl)-substituted GTP Analogs: A Novel Class of Potent Competitive Adenylyl Cyclase Inhibitors*

Andreas Gille and Roland SeifertDagger

From the Department of Pharmacology and Toxicology, the University of Kansas, Lawrence, Kansas 66045-7582

2'(3')-O-(N-Methylanthraniloyl)-(MANT)-substituted nucleotides are fluorescent and widely used for the kinetic analysis of enzymes and signaling proteins. We studied the effects of MANT-guanosine 5'-[gamma -thio]triphosphate (MANT-GTPgamma S) and MANT-guanosine 5'-[beta ,gamma -imido]triphosphate (MANT-GppNHp) on Galpha s- and Galpha i-protein-mediated signaling. MANT-GTPgamma S/MANT-GppNHp had lower affinities for Galpha s and Galpha i than GTPgamma S/GppNHp as assessed by inhibition of GTP hydrolysis of receptor-Galpha fusion proteins. MANT-GTPgamma S was much less effective than GTPgamma S at disrupting the ternary complex between the formyl peptide receptor and Galpha i2. MANT-GTPgamma S/MANT-GppNHp non-competitively inhibited GTPgamma S/GppNHp-, AlF<UP><SUB>4</SUB><SUP>−</SUP></UP>-, beta 2-adrenoceptor plus GTP-, cholera toxin plus GTP-, and forskolin-stimulated adenylyl cyclase (AC) in Galpha s-expressing Sf9 insect cell membranes and S49 wild-type lymphoma cell membranes. AC inhibition by MANT-GTPgamma S/MANT-GppNHp was not due to Galpha s inhibition because it was also observed in Galpha s-deficient S49 cyc- lymphoma cell membranes. Mn2+ blocked AC inhibition by GTPgamma S/GppNHp in S49 cyc- membranes but enhanced the potency of MANT-GTPgamma S/MANT-GppNHp at inhibiting AC by ~4-8-fold. MANT-GTPgamma S and MANT-GppNHp competitively inhibited forskolin/Mn2+-stimulated AC in S49 cyc- membranes with Ki values of 53 and 160 nM, respectively. The Ki value for MANT-GppNHp at insect cell AC was 155 nM. Collectively, MANT-GTPgamma S/MANT-GppNHp bind to Galpha s- and Galpha i-proteins with low affinity and are ineffective at activating Galpha . Instead, MANT-GTPgamma S/MANT-GppNHp constitute a novel class of potent competitive AC inhibitors.

* This work was supported by Army Research Office Grant DAAD19-00-1-0069, The J. R. and Inez Jay Biomedical Research Award of the University of Kansas (to R. S.), and a predoctoral fellowship from the Studienstiftung des Deutschen Volkes (to A. G.).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.

Dagger To whom correspondence should be addressed: Dept. of Pharmacology and Toxicology, the University of Kansas, 1251 Wescoe Hall Dr., Malott Hall, Rm. 5064, Lawrence, KS 66045-7582. Tel.: 785-864-3525; Fax: 785-864-5219; E-mail: rseifert@ku.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
 Mol. Pharmacol.Home page
M. Gottle, S. Dove, P. Steindel, Y. Shen, W.-J. Tang, J. Geduhn, B. Konig, and R. Seifert
Molecular Analysis of the Interaction of Bordetella pertussis Adenylyl Cyclase with Fluorescent Nucleotides
Mol. Pharmacol., September 1, 2007; 72(3): 526 - 535.
[Abstract] [Full Text] [PDF]

Home page
 J. Pharmacol. Exp. Ther.Home page
D. Rottlaender, J. Matthes, S. F. Vatner, R. Seifert, and S. Herzig
Functional Adenylyl Cyclase Inhibition in Murine Cardiomyocytes by 2'(3')-O-(N-Methylanthraniloyl)-Guanosine 5'-[{gamma}-Thio]triphosphate
J. Pharmacol. Exp. Ther., May 1, 2007; 321(2): 608 - 615.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Pharmacol.Home page
T.-C. Mou, A. Gille, S. Suryanarayana, M. Richter, R. Seifert, and S. R. Sprang
Broad Specificity of Mammalian Adenylyl Cyclase for Interaction with 2',3'-Substituted Purine- and Pyrimidine Nucleotide Inhibitors
Mol. Pharmacol., September 1, 2006; 70(3): 878 - 886.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T.-C. Mou, A. Gille, D. A. Fancy, R. Seifert, and S. R. Sprang
Structural Basis for the Inhibition of Mammalian Membrane Adenylyl Cyclase by 2 '(3')-O-(N-Methylanthraniloyl)-guanosine 5 '-Triphosphate
J. Biol. Chem., February 25, 2005; 280(8): 7253 - 7261.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. Gille, G. H. Lushington, T.-C. Mou, M. B. Doughty, R. A. Johnson, and R. Seifert
Differential Inhibition of Adenylyl Cyclase Isoforms and Soluble Guanylyl Cyclase by Purine and Pyrimidine Nucleotides
J. Biol. Chem., May 7, 2004; 279(19): 19955 - 19969.
[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.