Protein Motions Are Coupled to the Reaction Chemistry in Coenzyme B<sub>12</sub>‐Dependent Ethanolamine Ammonia Lyase

Russell, Henry J.; Jones, Alex R.; Hay, Sam; Greetham, Gregory M.; Towrie, Michael; Scrutton, Nigel S.

doi:10.1002/anie.201202502

Cited by 28 publications

(30 citation statements)

References 36 publications

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“…Similar findings have been found in other enzymes, especially those in which quantum tunneling may play a role . Quantum tunneling behavior is often probed using kinetic isotope effects (KIEs) (see reviews).…”

Section: Enzyme Structural Dynamics and The Chemical Stepsupporting

confidence: 63%

Engineered control of enzyme structural dynamics and function

2018

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Enzymes undergo a range of internal motions from local, active site fluctuations to large-scale, global conformational changes. These motions are often important for enzyme function, including in ligand binding and dissociation and even preparing the active site for chemical catalysis. Protein engineering efforts have been directed towards manipulating enzyme structural dynamics and conformational changes, including targeting specific amino acid interactions and creation of chimeric enzymes with new regulatory functions. Post-translational covalent modification can provide an additional level of enzyme control. These studies have not only provided insights into the functional role of protein motions, but they offer opportunities to create stimulus-responsive enzymes. These enzymes can be engineered to respond to a number of external stimuli, including light, pH, and the presence of novel allosteric modulators. Altogether, the ability to engineer and control enzyme structural dynamics can provide new tools for biotechnology and medicine.

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Section: Enzyme Structural Dynamics and The Chemical Stepsupporting

confidence: 63%

Engineered control of enzyme structural dynamics and function

2018

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“…Recombinant EAL from Salmonella enterica was prepared as described previously 19. 34 Holo-EAL was prepared freshly on the day of experiments by reconstituting EAL with excess coenzyme B 12 prior to removal of unbound B 12 by size-exclusion chromatography using a 10 DG desalting column (BioRad, Hemel Hempstead, UK). [D 2 ]EAL was prepared by treating 10–20 μ M of holo-EAL with 2 m M [D 4 ]EA for 10 min prior to separation of products and unreacted substrate with a 10 DG desalting column.…”

Section: Methodsmentioning

confidence: 99%

Probing Reversible Chemistry in Coenzyme B₁₂‐Dependent Ethanolamine Ammonia Lyase with Kinetic Isotope Effects

Jones

Rentergent

Scrutton

et al. 2015

Chemistry A European J

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Coenzyme B12-dependent enzymes such as ethanolamine ammonia lyase have remarkable catalytic power and some unique properties that enable detailed analysis of the reaction chemistry and associated dynamics. By selectively deuterating the substrate (ethanolamine) and/or the β-carbon of the 5′-deoxyadenosyl moiety of the intrinsic coenzyme B12, it was possible to experimentally probe both the forward and reverse hydrogen atom transfers between the 5′-deoxyadenosyl radical and substrate during single-turnover stopped-flow measurements. These data are interpreted within the context of a kinetic model where the 5′-deoxyadenosyl radical intermediate may be quasi-stable and rearrangement of the substrate radical is essentially irreversible. Global fitting of these data allows estimation of the intrinsic rate constants associated with CoC homolysis and initial H-abstraction steps. In contrast to previous stopped-flow studies, the apparent kinetic isotope effects are found to be relatively small.

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“…First, AdoCbl photolysis creates the same RP. The photophysical and photochemical dynamics of both unbound [17,18,[54][55][56][57][58][59] and enzyme-bound [60][61][62][63][64] AdoCbl have been well characterized, allowing us to accurately assess any perturbation of these dynamics from the protein. After excitation in water, there is rapid relaxation from the excited state(s) to a cob(II) alamin-like species (Fig.…”

Section: Coenzyme B 12 -Dependent Enzymology: a Physical Chemistry Pementioning

confidence: 99%

Relating localized protein motions to the reaction coordinate in coenzyme B₁₂‐dependent enzymes

et al. 2013

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The classical picture of enzyme catalysis relies on controlling the entropic and enthalpic contributions by manipulating reaction barriers and co-locating reactants and cofactors to facilitate the reaction chemistry. Catalysis is linked inextricably to the geometry of the enzyme-substrate complex and the chemical/physical properties of the active site, and probably to dynamical contributions that guide reactants along the desired reaction coordinate. Coenzyme B 12 -dependent enzymes have remarkable catalytic power and unique properties that enable detailed analysis of the reaction chemistry and associated dynamics. Here we discuss recent developments that are beginning to provide atomistic insight into how coenzyme B 12 -dependent enzymes steer reactants along the reaction coordinate. Such insight will ultimately generate 'movies' of the catalytic process across all relevant time scales. In the longer term, this will enable more predictive engineering of this class of enzyme to achieve new and desirable chemical outcomes.

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Protein Motions Are Coupled to the Reaction Chemistry in Coenzyme B₁₂‐Dependent Ethanolamine Ammonia Lyase

Cited by 28 publications

References 36 publications

Engineered control of enzyme structural dynamics and function

Engineered control of enzyme structural dynamics and function

Probing Reversible Chemistry in Coenzyme B₁₂‐Dependent Ethanolamine Ammonia Lyase with Kinetic Isotope Effects

Relating localized protein motions to the reaction coordinate in coenzyme B₁₂‐dependent enzymes

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Protein Motions Are Coupled to the Reaction Chemistry in Coenzyme B12‐Dependent Ethanolamine Ammonia Lyase

Cited by 28 publications

References 36 publications

Engineered control of enzyme structural dynamics and function

Engineered control of enzyme structural dynamics and function

Probing Reversible Chemistry in Coenzyme B12‐Dependent Ethanolamine Ammonia Lyase with Kinetic Isotope Effects

Relating localized protein motions to the reaction coordinate in coenzyme B12‐dependent enzymes

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Protein Motions Are Coupled to the Reaction Chemistry in Coenzyme B₁₂‐Dependent Ethanolamine Ammonia Lyase

Probing Reversible Chemistry in Coenzyme B₁₂‐Dependent Ethanolamine Ammonia Lyase with Kinetic Isotope Effects

Relating localized protein motions to the reaction coordinate in coenzyme B₁₂‐dependent enzymes