HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Brundiers, R. Articles by Konrad, M. Search for Related Content PubMed PubMed Citation Articles by Brundiers, R. Articles by Konrad, M.

J Biol Chem, Vol. 274, Issue 50, 35289-35292, December 10, 1999

COMMUNICATION
Modifying Human Thymidylate Kinase to Potentiate Azidothymidine Activation^*

Ralf Brundiers, Arnon Lavie^§, Thomas Veit, Jochen Reinstein, Ilme Schlichting^¶, Nils Ostermann, Roger S. Goody, and Manfred Konrad^**

From the Department of Molecular Genetics, Max Planck Institute for Biophysical Chemistry, D-37070 Göttingen, Germany and the  Department of Physical Biochemistry, Max Planck Institute for Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund, Germany

Based on the knowledge of the crystal structures of yeast and Escherichia coli thymidylate kinases (TmpKs) and the observation that TmpK from E. coli can phosphorylate azidothymidine monophosphate (AZT-MP) much more efficiently than either the yeast or the highly homologous human enzyme, we have engineered yeast and human TmpKs to obtain enzymes that have dramatically improved AZT-MP phosphorylation properties. These modified enzymes have properties that make them attractive candidates for gene therapeutic approaches to potentiating the action of AZT as an inhibitor of human immunodeficiency virus (HIV) replication. In particular, insertion of the lid domain of the bacterial TmpK into the human enzyme results in a pronounced change of the acceptance of AZT-MP such that it is now phosphorylated even faster than TMP.

---

^* 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.

^§ Supported by the Peter and Traudl Engelhorn Stiftung.

^¶ Supported by the Richard and Anne-Liese Gielen-Leyendecker Stiftung.

To whom correspondence may be addressed. E-mail: roger. goody@mpi-dortmund.mpg.de.

^** To whom correspondence may also be addressed. E-mail: mkonrad@gwdg.de.

---

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.

This article has been cited by other articles:

				N. Sekulic, M. Konrad, and A. Lavie Structural Mechanism for Substrate Inhibition of the Adenosine 5'-Phosphosulfate Kinase Domain of Human 3'-Phosphoadenosine 5'-Phosphosulfate Synthetase 1 and Its Ramifications for Enzyme Regulation J. Biol. Chem., July 27, 2007; 282(30): 22112 - 22121. [Abstract] [Full Text] [PDF]

				C.-H. Hsu, R. Hu, G. E. Dutschman, G. Yang, P. Krishnan, H. Tanaka, M. Baba, and Y.-C. Cheng Comparison of the Phosphorylation of 4'-Ethynyl 2',3'-Dihydro-3'-Deoxythymidine with That of Other Anti-Human Immunodeficiency Virus Thymidine Analogs Antimicrob. Agents Chemother., May 1, 2007; 51(5): 1687 - 1693. [Abstract] [Full Text] [PDF]

				D.-N. Chaperon Construction and Complementation of In-Frame Deletions of the Essential Escherichia coli Thymidylate Kinase Gene Appl. Envir. Microbiol., February 1, 2006; 72(2): 1288 - 1294. [Abstract] [Full Text] [PDF]

				B. M. Wohrl, L. Loubiere, R. Brundiers, R. S. Goody, D. Klatzmann, and M. Konrad Expressing engineered thymidylate kinase variants in human cells to improve AZT phosphorylation and human immunodeficiency virus inhibition J. Gen. Virol., March 1, 2005; 86(3): 757 - 764. [Abstract] [Full Text] [PDF]

				D. Segura-Pena, N. Sekulic, S. Ort, M. Konrad, and A. Lavie Substrate-induced Conformational Changes in Human UMP/CMP Kinase J. Biol. Chem., August 6, 2004; 279(32): 33882 - 33889. [Abstract] [Full Text] [PDF]

				I. Paarmann, O. Spangenberg, A. Lavie, and M. Konrad Formation of Complexes between Ca2+{middle dot}Calmodulin and the Synapse-associated Protein SAP97 Requires the SH3 Domain-Guanylate Kinase Domain-connecting HOOK Region J. Biol. Chem., October 18, 2002; 277(43): 40832 - 40838. [Abstract] [Full Text] [PDF]

				S. Gallois-Montbrun, B. Schneider, Y. Chen, V. Giacomoni-Fernandes, L. Mulard, S. Morera, J. Janin, D. Deville-Bonne, and M. Veron Improving Nucleoside Diphosphate Kinase for Antiviral Nucleotide Analogs Activation J. Biol. Chem., October 11, 2002; 277(42): 39953 - 39959. [Abstract] [Full Text] [PDF]

				N. Sekulic, L. Shuvalova, O. Spangenberg, M. Konrad, and A. Lavie Structural Characterization of the Closed Conformation of Mouse Guanylate Kinase J. Biol. Chem., August 9, 2002; 277(33): 30236 - 30243. [Abstract] [Full Text] [PDF]

				Y. Li, O. Spangenberg, I. Paarmann, M. Konrad, and A. Lavie Structural Basis for Nucleotide-dependent Regulation of Membrane-associated Guanylate Kinase-like Domains J. Biol. Chem., February 1, 2002; 277(6): 4159 - 4165. [Abstract] [Full Text] [PDF]