Mechanism of Thermal Adaptation in the Lactate Dehydrogenases

Huo Lei Peng, Tsuyoshi Egawa, Eric Chang, Hua Deng, Robert Callender

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The mechanism of thermal adaptation of enzyme function at the molecular level is poorly understood but is thought to lie within the structure of the protein or its dynamics. Our previous work on pig heart lactate dehydrogenase (phLDH) has determined very high resolution structures of the active site, via isotope edited IR studies, and has characterized its dynamical nature, via laser-induced temperature jump (T-jump) relaxation spectroscopy on the Michaelis complex. These particular probes are quite powerful at getting at the interplay between structure and dynamics in adaptation. Hence, we extend these studies to the psychrophilic protein cgLDH (Champsocephalus gunnari; 0 °C) and the extreme thermophile tmLDH (Thermotoga maritima LDH; 80 °C) for comparison to the mesophile phLDH (38-39 °C). Instead of the native substrate pyruvate, we utilize oxamate as a nonreactive substrate mimic for experimental reasons. Using isotope edited IR spectroscopy, we find small differences in the substate composition that arise from the detailed bonding patterns of oxamate within the active site of the three proteins; however, we find these differences insufficient to explain the mechanism of thermal adaptation. On the other hand, T-jump studies of reduced β-nicotinamide adenine dinucleotide (NADH) emission reveal that the most important parameter affecting thermal adaptation appears to be enzyme control of the specific kinetics and dynamics of protein motions that lie along the catalytic pathway. The relaxation rate of the motions scale as cgLDH > phLDH > tmLDH in a way that faithfully matches kcat of the three isozymes.

Original languageEnglish (US)
Pages (from-to)15256-15262
Number of pages7
JournalJournal of Physical Chemistry B
Volume119
Issue number49
DOIs
StatePublished - Dec 10 2015

Fingerprint

Lactate Dehydrogenases
lactates
dehydrogenases
swine
L-Lactate Dehydrogenase
proteins
Proteins
Isotopes
NAD
enzymes
mesophiles
thermophiles
Enzymes
isotopes
pyruvates
nicotinamide
Enzyme kinetics
adenines
Substrates
Pyruvic Acid

ASJC Scopus subject areas

  • Physical and Theoretical Chemistry
  • Materials Chemistry
  • Surfaces, Coatings and Films

Cite this

Mechanism of Thermal Adaptation in the Lactate Dehydrogenases. / Peng, Huo Lei; Egawa, Tsuyoshi; Chang, Eric; Deng, Hua; Callender, Robert.

In: Journal of Physical Chemistry B, Vol. 119, No. 49, 10.12.2015, p. 15256-15262.

Research output: Contribution to journalArticle

Peng, Huo Lei ; Egawa, Tsuyoshi ; Chang, Eric ; Deng, Hua ; Callender, Robert. / Mechanism of Thermal Adaptation in the Lactate Dehydrogenases. In: Journal of Physical Chemistry B. 2015 ; Vol. 119, No. 49. pp. 15256-15262.
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