Browsing QFA_Articles by Author "c1e8643d-ab2b-4197-aa22-d33001db8e80"
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Chemical Ligation and Isotope Labeling to Locate Dynamic Effects during Catalysis by Dihydrofolate Reductase
Luk, Louis Y. P.; Ruiz-Pernía, José Javier; Adesina, Aduragbemi S.; Loveridge, E. Joel; Tuñón, Iñaki; Moliner, Vicent; Allemann, Rudolf K. Wiley (2015-07-27)Experimental and computational approaches have long been employed to define the role of protein motions in en- zyme catalysis, but a refined experimental method for locating the origin of dynamic effects has not previously ... -
Heavy Enzymes and the Rational Redesign of Protein Catalysts
Scott, Alan; Luk, Louis Y. P.; Tuñón, Iñaki; Moliner, Vicent; Allemann, Rudolf K. Wiley (2019-11-18)An unsolved mystery in biology concerns the link between enzyme catalysis and protein motions. Comparison between isotopically labelled “heavy” dihydrofolate reductases and their natural‐abundance counterparts has suggested ... -
Increased Dynamic Effects in a Catalytically Compromised Variant of Escherichia coli Dihydrofolate Reductase
Ruiz-Pernía, José Javier; Luk, Louis Y. P.; García Meseguer, Rafael; Martí Forés, Sergio; Loveridge, E. Joel; Tuñón, Iñaki; Moliner, Vicent; Allemann, Rudolf K. American Chemical Society (2013)Isotopic substitution (15N, 13C, 2H) of a catalytically compromised variant of Escherichia coli dihydrofolate reductase, EcDHFR-N23PP/S148A, has been used to investigate the effect of these mutations on catalysis. The ... -
Isotope Substitution of Promiscuous Alcohol Dehydrogenase Reveals the Origin of Substrate Preference in the Transition State
Behiry, Enas; Ruiz-Pernía, José Javier; Luk, Louis Y. P.; Tuñón, Iñaki; Moliner, Vicent; Allemann, Rudolf K. Wiley (2018-03-12)The origin of substrate preference in promiscuous enzymes was investigated by enzyme isotope labelling of the alcohol dehydrogenase from Geobacillus stearothermophilus (BsADH). At physiological temperature, protein dynamic ... -
Loss of Hyperconjugative Effects Drives Hydride Transfer during Dihydrofolate Reductase Catalysis
Angelastro, Antonio; Ruiz-Pernía, José Javier; Tuñón, Iñaki; Moliner, Vicent; Luk, Louis Y. P.; Allemann, Rudolf K. American Chemical Society (2019-09-23)Hydride transfer is widespread in nature and has an essential role in applied research. However, the mechanisms of how this transformation occurs in living organisms remain a matter of vigorous debate. Here, we examined ... -
Minimization of dynamic effects in the evolution of dihydrofolate reductase
Ruiz-Pernía, José Javier; Behiry, Enas; Luk, Louis Y. P.; Loveridge, E. Joel; Tuñón, Iñaki; Moliner, Vicent; Allemann, Rudolf K. Royal Society of Chemistry (2016)Protein isotope labeling is a powerful technique to probe functionally important motions in enzyme catalysis and can be applied to investigate the conformational dynamics of proteins. Previous investigations have indicated ... -
Protein Isotope Effects in Dihydrofolate Reductase From Geobacillus stearothermophilus Show Entropic–Enthalpic Compensatory Effects on the Rate Constant
Luk, Louis Y. P.; Ruiz-Pernía, José Javier; Dawson, William M.; Loveridge, E. Joel; Tuñón, Iñaki; Moliner, Vicent; Allemann, Rudolf K. American Chemical Society (2014)Catalysis by dihydrofolate reductase from the moderately thermophilic bacterium Geobacillus stearothermophilus (BsDHFR) was investigated by isotope substitution of the enzyme. The enzyme kinetic isotope effect for hydride ... -
Unraveling the role of protein dynamics in dihydrofolate reductase catalysis
Moliner, Vicent; Ruiz-Pernía, José Javier; Luk, Louis Y. P.; Dawson, William M.; Roca, Maite; Loveridge, E. Joel; Glowacki, David R.; Harvey, Jeremy N.; Mulholland, Adrian J.; Tuñón, Iñaki; Allemann, Rudolf K. National Academy of Sciences (2013)Protein dynamics have controversially been proposed to be at the heart of enzyme catalysis, but identification and analysis of dynamical effects in enzyme-catalyzed reactions have proved very challenging. Here, we tackle ... -
Why Are Some Enzymes Dimers? Flexibility and Catalysis in Thermotoga maritima Dihydrofolate Reductase
Ruiz-Pernía, José Javier; Tuñón, Iñaki; Moliner, Vicent; Allemann, Rudolf K. American Chemical Society (2019-05-13)Dihydrofolate reductase from Thermotoga maritima (TmDFHFR) is a dimeric thermophilic enzyme that catalyzes the hydride transfer from the cofactor NADPH to dihydrofolate less efficiently than other DHFR enzymes, such as the ...