Mapping the Accessible Conformational Landscape of an Insect Carboxylesterase Using Conformational Ensemble Analysis and Kinetic Crystallography

Galen J. Correy, Paul D. Carr, Tamara Meirelles, Peter D. Mabbitt, Nicholas J. Fraser, Martin Weik, Colin J. Jackson (Lead / Corresponding author)

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

The proper function of enzymes often depends upon their efficient interconversion between particular conformational sub-states on a free-energy landscape. Experimentally characterizing these sub-states is challenging, which has limited our understanding of the role of protein dynamics in many enzymes. Here, we have used a combination of kinetic crystallography and detailed analysis of crystallographic protein ensembles to map the accessible conformational landscape of an insect carboxylesterase (LcαE7) as it traverses all steps in its catalytic cycle. LcαE7 is of special interest because of its evolving role in organophosphate insecticide resistance. Our results reveal that a dynamically coupled network of residues extends from the substrate-binding site to a surface loop. Interestingly, the coupling of this network that is apparent in the apoenzyme appears to be reduced in the phosphorylated enzyme intermediate. Altogether, the results of this work highlight the importance of protein dynamics to enzyme function and the evolution of new activity.

Original languageEnglish
Pages (from-to)977-87
Number of pages11
JournalStructure
Volume24
Issue number6
Early online date19 May 2016
DOIs
Publication statusPublished - 7 Jun 2016

Keywords

  • Carboxylesterase/chemistry
  • Crystallography, X-Ray
  • Insect Proteins/chemistry
  • Kinetics
  • Models, Molecular
  • Protein Binding
  • Protein Conformation

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