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Originally published In Press as doi:10.1074/jbc.M011396200 on March 19, 2001

J. Biol. Chem., Vol. 276, Issue 21, 18090-18095, May 25, 2001
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Identification of New JNK Substrate Using ATP Pocket Mutant JNK and a Corresponding ATP Analogue*

Hasem Habelhah, Kavita ShahDagger , Lan Huang§, Alma L. Burlingame§, Kevan M. ShokatDagger , and Ze'ev Ronai

From the Ruttenberg Cancer Center, Mount Sinai School of Medicine, New York, New York 10029 and the Dagger  Department of Cellular and Molecular Pharmacology, and § Pharmaceutical Chemistry, University of California, San Francisco, California 94143

Modification of the ATP pocket on protein kinases allows selective use of an ATP analogue that exhibits high affinity for the altered kinases. Using this approach, we altered the ATP-binding site on JNK and identified N6-(2-phenythyl)-ATP, a modified form of ATP that exhibits high specificity and affinity for the modified, but not the wild type form, of JNK. Using modified JNK and its ATP analogue enables the detection of novel JNK substrates. Among substrates identified using this approach is heterogeneous nuclear ribonucleoprotein K, which is involved in transcription and post-transcriptional mRNA metabolism. The newly identified substrate can be phosphorylated by JNK on amino acids 216 and 353, which contribute to heterogeneous nuclear ribonucleoprotein K mediated transcriptional activities.

* This work was supported by National Institutes of Health Grants CA59008 (to Z. R.), CA70731 (to K. M. S.), and RR01614 (to A. L. B.).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: Ruttenberg Cancer Center, Mount Sinai School of Medicine, 1425 Madison Ave., Rm. 15-20, New York, NY 10029. Fax: 212-849-2425; E-mail: zeev. ronai{at}mssm.edu.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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