Global conformational changes in allosteric proteins: A study of Escherichia coli cAMP receptor protein and muscle pyruvate kinase

Ewa Heyduk, Tomasz Heyduk, James Lee

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

One of the basic features in allosteric regulation involves long range transduction of information. Based on crystallographic data on protein systems that are regulated by allosteric mechanisms, a global conformational change has always been observed. It is, therefore, important and useful to correlate the cooperativity of global structural change with the mode of binding of the regulatory ligand. Two systems were chosen for study, namely Escherichia coli cAMP receptor protein and muscle pyruvate kinase, which show negative and positive cooperativity in the binding of allosteric ligands, respectively. Quantitative titration of the global structural change, monitored by a high precision analytical gel chromatography technique, was conducted as a function of allosteric effector concentration. The results obtained for cAMP receptor protein show that the protein undergoes contraction upon binding of cAMP. The decreases in Stokes radius associated with complex formation are 0.1 ± 0.1 and 0.7 ± 0.1 Å when one and two cAMP-binding sites are filled, respectively. The results for the pyruvate kinase system show a concerted structural change that quantitatively match the predicted behavior based on equilibrium constants derived from the analysis of steady state kinetic data by a two-state model. Hence, for these two systems, these results show that negative and positive cooperativity are correlated with sequential and concerted modes of structural change, respectively.

Original languageEnglish (US)
Pages (from-to)3200-3204
Number of pages5
JournalJournal of Biological Chemistry
Volume267
Issue number5
StatePublished - Feb 15 1992

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Cyclic AMP Receptor Protein
Pyruvate Kinase
Escherichia coli Proteins
Staphylococcal Protein A
Escherichia coli
Muscle
Allosteric Regulation
Ligands
Muscles
Equilibrium constants
Chromatography
Titration
Gel Chromatography
Proteins
Gels
Binding Sites
Kinetics

ASJC Scopus subject areas

  • Biochemistry

Cite this

Global conformational changes in allosteric proteins : A study of Escherichia coli cAMP receptor protein and muscle pyruvate kinase. / Heyduk, Ewa; Heyduk, Tomasz; Lee, James.

In: Journal of Biological Chemistry, Vol. 267, No. 5, 15.02.1992, p. 3200-3204.

Research output: Contribution to journalArticle

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