Domain interaction in rabbit muscle pyruvate kinase. I. Effects of ligands on protein denaturation induced by guanidine hydrochloride

T. G. Consler, James Lee

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

The structural stability of rabbit muscle pyruvate kinase was examined. The unfolding of pyruvate kinase was induced by guanidine hydrochloride, and the process was monitored by spectroscopic techniques (fluorescence and UV absorption) and hydrodynamic measurements (sedimentation velocity, sedimentation equilibrium, densimetry, and viscometry). The spectroscopic techniques revealed that the unfolding of pyruvate kinase induced by guanidine hydrochloride is not a simple cooperative process. This suggests that different regions of pyruvate kinase are unfolding with different efficiencies in response to the denaturant. These regions are most likely related to the domain structures observed by x-ray crystallography. In the presence of L-phenylalanine, the allosteric inhibitor, the denaturation process became more cooperative, and the enzyme dissociated and unfolded at a higher denaturant concentration. The binding of phenylalanine also induced a structural change in the enzyme, rendering it more susceptible to tryptic digestion. One of the peptides, the production rate of which was increased, was isolated and sequenced. Its N terminus is located at the interface between two domains, one of which contains the active site. This evidence indicates structural changes, probably involving domain-domain interaction, for pyruvate kinase in response to phenylalanine binding.

Original languageEnglish (US)
Pages (from-to)2787-2793
Number of pages7
JournalJournal of Biological Chemistry
Volume263
Issue number6
StatePublished - 1988
Externally publishedYes

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Protein Denaturation
Denaturation
Pyruvate Kinase
Guanidine
Muscle
Rabbits
Ligands
Muscles
Phenylalanine
Proteins
Sedimentation
Crystallography
Viscosity measurement
Enzymes
Hydrodynamics
Digestion
Catalytic Domain
Fluorescence
X-Rays
X rays

ASJC Scopus subject areas

  • Biochemistry

Cite this

Domain interaction in rabbit muscle pyruvate kinase. I. Effects of ligands on protein denaturation induced by guanidine hydrochloride. / Consler, T. G.; Lee, James.

In: Journal of Biological Chemistry, Vol. 263, No. 6, 1988, p. 2787-2793.

Research output: Contribution to journalArticle

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