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  Insights into the phosphoryltransfer mechanism of human thymidylate kinase gained from crystal structures of enzyme complexes along the reaction coordinate

Ostermann, N., Schlichting, I., Brundiers, R., Konrad, M., Reinstein, J., Veit, T., et al. (2000). Insights into the phosphoryltransfer mechanism of human thymidylate kinase gained from crystal structures of enzyme complexes along the reaction coordinate. Structure, 8(6), 629-642. doi:10.1016/S0969-2126(00)00149-0.

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Genre: Journal Article
Alternative Title : Insights into the phosphoryltransfer mechanism of human thymidylate kinase gained from crystal structures of enzyme complexes along the reaction coordinate

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 Creators:
Ostermann, Nils, Author
Schlichting, Ilme1, Author              
Brundiers, Ralf, Author
Konrad, Manfred, Author
Reinstein, Jochen1, Author              
Veit, Thomas, Author
Goody, Roger S.2, Author              
Lavie, Amon, Author
Affiliations:
1Department of Biomolecular Mechanisms, Max Planck Institute for Medical Research, Max Planck Society, ou_1497700              
2Emeritus Group Biophysics, Max Planck Institute for Medical Research, Max Planck Society, ou_1497712              

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Free keywords: Conformational change; Crystal structures; P loop; Thymidylate kinase
 Abstract: Background: Thymidylate kinase (TMPK) is a nucleoside monophosphate kinase that catalyzes the reversible phosphoryltransfer between ATP and TMP to yield ADP and TDP. In addition to its vital role in supplying precursors for DNA synthesis, human TMPK has an important medical role participating in the activation of a number of anti-HIV prodrugs. Results: Crystal structures of human TMPK in complex with TMP and ADP, TMP and the ATP analog AppNHp, TMP with ADP and the phosphoryl analog AlF3, TDP and ADP, and the bisubstrate analog TP5A were determined. The conformations of the P-loop, the LID region, and the adenine-binding loop vary according to the nature of the complex. Substitution of ADP by AppNHp results in partial closure of the P-loop and the rotation of the TMP phosphate group to a catalytically unfavorable position, which rotates back in the AlF3 complex to a position suitable for in-line attack. In the fully closed state observed in the TP5A and the TDP–ADP complexes, Asp15 interacts strongly with the 3′-hydroxyl group of TMP. Conclusions: The observed changes of nucleotide state and conformation and the corresponding protein structural changes are correlated with intermediates occurring along the reaction coordinate and show the sequence of events occurring during phosphate transfer. The low catalytic activity of human TMPK appears to be determined by structural changes required to achieve catalytic competence and it is suggested that a mechanism might exist to accelerate the activity.

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Language(s): eng - English
 Dates: 2000-03-072000-02-102000-03-232000-06-162000-06-15
 Publication Status: Published in print
 Pages: 14
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 666305
DOI: 10.1016/S0969-2126(00)00149-0
Other: 4808
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Title: Structure
  Other : Structure
Source Genre: Journal
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Publ. Info: London : Cell Press
Pages: - Volume / Issue: 8 (6) Sequence Number: - Start / End Page: 629 - 642 Identifier: ISSN: 0969-2126
CoNE: https://pure.mpg.de/cone/journals/resource/954927002244_1