Effects of metabolites on the structural dynamics of rabbit muscle pyruvate kinase

Shaoning Yu, Lucy L Y Lee, James Lee

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

The activity of rabbit muscle pyruvate kinase (PK) is regulated by metabolites. Besides requiring the presence of its substrates, PEP and ADP, the enzyme requires Mg2+ and K+ for activity. PK is allosterically inhibited by Phe for activity. The presence of PEP or Phe has opposing effects on the hydrodynamic properties of the enzyme without an apparent change in secondary structure. In this study, the structural perturbation induced by ligand binding was investigated by Fourier transform infrared (FT-IR) spectroscopy. Furthermore, the structural dynamics of PK was probed by H/D exchange monitored by FT-IR. Substrates and activating metal ions induce PK to assume a more dynamic structure while Phe exerts an opposite effect. In all cases there is no significant interconversion of secondary structures. PEP is the most efficient ligand in inducing a change in the microenvironments of both helices and sheets so much so that they can be detected spectroscopically as separate bands. These results provide the first evidence for a differential effect of ligand binding on the dynamics of structural elements in PK. Furthermore, the data support the model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the observed change in hydrodynamic properties represent the two extreme end states.

Original languageEnglish (US)
Pages (from-to)1-11
Number of pages11
JournalBiophysical Chemistry
Volume103
Issue number1
DOIs
StatePublished - Jan 8 2003

Fingerprint

pyruvates
Pyruvate Kinase
dynamic structural analysis
metabolites
rabbits
Structural dynamics
Metabolites
muscles
Muscle
Rabbits
Muscles
Hydrodynamics
Ligands
ligands
enzymes
hydrodynamics
Allosteric Regulation
adenosine diphosphate
perturbation
Fourier Analysis

Keywords

  • Allostery
  • Dynamics
  • Fourier transform infrared spectroscopy
  • Pyruvate kinase

ASJC Scopus subject areas

  • Biochemistry
  • Physical and Theoretical Chemistry
  • Biophysics

Cite this

Effects of metabolites on the structural dynamics of rabbit muscle pyruvate kinase. / Yu, Shaoning; Lee, Lucy L Y; Lee, James.

In: Biophysical Chemistry, Vol. 103, No. 1, 08.01.2003, p. 1-11.

Research output: Contribution to journalArticle

Yu, Shaoning ; Lee, Lucy L Y ; Lee, James. / Effects of metabolites on the structural dynamics of rabbit muscle pyruvate kinase. In: Biophysical Chemistry. 2003 ; Vol. 103, No. 1. pp. 1-11.
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