The energy landscape of adenylate kinase during catalysis

S. Jordan Kerns, Roman V. Agafonov, Young Jin Cho, Francesco Pontiggia, Renee Otten, Dimitar V. Pachov, Steffen Kutter, Lien A. Phung, Padraig N. Murphy, Vu Thai, Tom Alber, Michael F. Hagan, Dorothee Kern

Research output: Contribution to journalArticlepeer-review

137 Scopus citations

Abstract

Kinases perform phosphoryl-transfer reactions in milliseconds; without enzymes, these reactions would take about 8,000 years under physiological conditions. Despite extensive studies, a comprehensive understanding of kinase energy landscapes, including both chemical and conformational steps, is lacking. Here we scrutinize the microscopic steps in the catalytic cycle of adenylate kinase, through a combination of NMR measurements during catalysis, pre-steady-state kinetics, molecular-dynamics simulations and crystallography of active complexes. We find that the Mg2+ cofactor activates two distinct molecular events: phosphoryl transfer (>105-fold) and lid opening (103-fold). In contrast, mutation of an essential active site arginine decelerates phosphoryl transfer 103-fold without substantially affecting lid opening. Our results highlight the importance of the entire energy landscape in catalysis and suggest that adenylate kinases have evolved to activate key processes simultaneously by precise placement of a single, charged and very abundant cofactor in a preorganized active site.

Original languageEnglish (US)
Pages (from-to)124-131
Number of pages8
JournalNature Structural and Molecular Biology
Volume22
Issue number2
DOIs
StatePublished - Jan 1 2015

Bibliographical note

Publisher Copyright:
© 2015 Nature America, Inc. All rights reserved.

Fingerprint

Dive into the research topics of 'The energy landscape of adenylate kinase during catalysis'. Together they form a unique fingerprint.

Cite this