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Unraveling Entropic Rate Acceleration Induced by Solvent Dynamics in Membrane Enzymes
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- Kürten, Charlotte (author)
- KTH,Science for Life Laboratory, SciLifeLab,Proteomik och nanobioteknologi
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- Syren, Per-Olof (author)
- KTH,Science for Life Laboratory, SciLifeLab,Proteomik och nanobioteknologi
- (creator_code:org_t)
- 2016-01-16
- 2016
- English.
- In: Journal of Visualized Experiments. - : Journal of Visualized Experiments. - 1940-087X. ; :107
- Journal article (peer-reviewed)
Abstract
Subject headings
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- Enzyme catalysis evolved in an aqueous environment. The influence of solvent dynamics on catalysis is, however, currently poorly understood and usually neglected. The study of water dynamics in enzymes and the associated thermodynamical consequences is highly complex and has involved computer simulations, nuclear magnetic resonance (NMR) experiments, and calorimetry. Water tunnels that connect the active site with the surrounding solvent are key to solvent displacement and dynamics. The protocol herein allows for the engineering of these motifs for water transport, which affects specificity, activity and thermodynamics. By providing a biophysical framework founded on theory and experiments, the method presented herein can be used by researchers without previous expertise in computer modeling or biophysical chemistry. The method will advance our understanding of enzyme catalysis on the molecular level by measuring the enthalpic and entropic changes associated with catalysis by enzyme variants with obstructed water tunnels. The protocol can be used for the study of membrane-bound enzymes and other complex systems. This will enhance our understanding of the importance of solvent reorganization in catalysis as well as provide new catalytic strategies in protein design and engineering.
Keyword
- Chemistry
- Issue 107
- Enzyme catalysis
- thermodynamics
- water
- dynamics
- membrane protein
- kinetics
- transition state theory
- protein engineering
- hydrophobic effect
Publication and Content Type
- ref (subject category)
- art (subject category)
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