Conformational heterogeneity within the Michaelis complex of lactate dehydrogenase

Hua Deng, Dung V. Vu, Keith Clinch, Ruel Desamero, R. Brian Dyer, Robert Callender

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

A series of isotope edited IR measurements, both static as well as temperature jump relaxation spectroscopy, are performed on lactate dehydrogenase (LDH) to determine the ensemble of structures available to its Michaelis complex. There clearly has been a substantial reduction in the number of states available to the pyruvate substrate (as modeled by the substrate mimic, oxamate) and NADH when bound to protein compared to dissolved in solution, as determined by the bandwidths and positions of the critical C2=O band of the bound substrate mimic and the C4-H stretch of the NADH reduced nicotinamide group. Moreover, it is found that a strong ionic bond (characterized by a signature IR band discovered in this study) is formed between the carboxyl group of bound pyruvate with (presumably) Arg171, forming a strong "anchor" within the protein matrix. However, conformational heterogeneity within the Michaelis complex is found that has an impact on both catalytic efficiency and thermodynamics of the enzyme.

Original languageEnglish (US)
Pages (from-to)7670-7678
Number of pages9
JournalJournal of Physical Chemistry B
Volume115
Issue number23
DOIs
StatePublished - Jun 16 2011

Fingerprint

lactates
dehydrogenases
L-Lactate Dehydrogenase
pyruvates
Pyruvic Acid
NAD
Substrates
proteins
Proteins
nicotinamide
Niacinamide
carboxyl group
Anchors
Isotopes
enzymes
Enzymes
isotopes
signatures
Spectroscopy
Thermodynamics

ASJC Scopus subject areas

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

Cite this

Conformational heterogeneity within the Michaelis complex of lactate dehydrogenase. / Deng, Hua; Vu, Dung V.; Clinch, Keith; Desamero, Ruel; Dyer, R. Brian; Callender, Robert.

In: Journal of Physical Chemistry B, Vol. 115, No. 23, 16.06.2011, p. 7670-7678.

Research output: Contribution to journalArticle

Deng, Hua ; Vu, Dung V. ; Clinch, Keith ; Desamero, Ruel ; Dyer, R. Brian ; Callender, Robert. / Conformational heterogeneity within the Michaelis complex of lactate dehydrogenase. In: Journal of Physical Chemistry B. 2011 ; Vol. 115, No. 23. pp. 7670-7678.
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