JBC

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


     

Originally published In Press as doi:10.1074/jbc.M003524200 on June 26, 2000

J. Biol. Chem., Vol. 275, Issue 39, 30394-30398, September 29, 2000
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
275/39/30394    most recent
M003524200v1
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 Ablooglu, A. J.
Right arrow Articles by Kohanski, R. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ablooglu, A. J.
Right arrow Articles by Kohanski, R. 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?

Probing the Catalytic Mechanism of the Insulin Receptor Kinase with a Tetrafluorotyrosine-containing Peptide Substrate*

Ararat J. AbloogluDagger §, Jeffrey H. Till§, Kyonghee Kim||, Keykavous Parang||, Philip A. Cole||, Stevan R. Hubbard, and Ronald A. KohanskiDagger **

From the Dagger  Mount Sinai School of Medicine, Department of Biochemistry and Molecular Biology, New York, New York, 10029  Skirball Institute of Biomolecular Medicine and Department of Pharmacology, New York University School of Medicine, New York, New York 10016, and || The Johns Hopkins University School of Medicine, Department of Pharmacology and Molecular Sciences, Baltimore, Maryland 21205

The interaction of a synthetic tetrafluorotyrosyl peptide substrate with the activated tyrosine kinase domain of the insulin receptor was studied by steady-state kinetics and x-ray crystallography. The pH-rate profiles indicate that the neutral phenol, rather than the chemically more reactive phenoxide ion, is required for enzyme-catalyzed phosphorylation. The pKa of the tetrafluorotyrosyl hydroxyl is elevated 2 pH units on the enzyme compared with solution, whereas the phenoxide anion species behaves as a weak competitive inhibitor of the tyrosine kinase. A structure of the binary enzyme-substrate complex shows the tetrafluorotyrosyl OH group at hydrogen bonding distances from the side chains of Asp1132 and Arg1136, consistent with elevation of the pKa. These findings strongly support a reaction mechanism favoring a dissociative transition state.

* This work was supported by Grant DK50074 from the National Institutes of Health (NIH) (to R. A. K.), Grant DK52916 from NIH (to S. R. H.), by the Winston Foundation (to K. K.), and by the Burroughs Welcome Fund and NIH Grant CA74305 (to P. A. C.).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.

§ Equal contributions were made by these authors.

** To whom correspondence should be addressed. Tel.: 212-241-7288; Fax: 212-996-7214; E-mail: Ronald.Kohanski@mssm.edu.

Copyright © 2000 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
K.-Y. Cheng, M. E. M. Noble, V. Skamnaki, N. R. Brown, E. D. Lowe, L. Kontogiannis, K. Shen, P. A. Cole, G. Siligardi, and L. N. Johnson
The Role of the Phospho-CDK2/Cyclin A Recruitment Site in Substrate Recognition
J. Biol. Chem., August 11, 2006; 281(32): 23167 - 23179.
[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 © 2000 by the American Society for Biochemistry and Molecular Biology.