Characterisation of a GroEL single-ring mutant that supports growth of Escherichia coli and has GroES-dependent ATPase activity

E Kovács, Z Sun, H Liu, DJ Scott, AI Karsisiotis, AR Clarke, SG Burston, PA Lund

Research output: Contribution to journalArticle (Academic Journal)

20 Citations (Scopus)

Abstract

Binding and folding of substrate proteins by the molecular chaperone GroEL alternates between its two seven-membered rings in an ATPregulated manner. The association of ATP and GroES to a polypeptidebound ring of GroEL encapsulates the folding proteins in the central cavity of that ring (cis ring) and allows it to fold in a protected environment where the risk of aggregation is reduced. ATP hydrolysis in the cis ring changes the potentials within the system such that ATP binding to the opposite (trans) ring triggers the release of all ligands from the cis ring of GroEL through a complex network of allosteric communication between the rings. Inter-ring allosteric communication thus appears indispensable for the function of GroEL, and an engineered single-ring version (SR1) cannot substitute for GroEL in vivo. We describe here the isolation and characterisation of an active single-ring form of the GroEL protein (SR-A92T), which has an exceptionally low ATPase activity that is strongly stimulated by the addition of GroES. Dissection of the kinetic pathway of the ATP-induced structural changes in this active single ring can be explained by the fact that the mutation effectively blocks progression through the full allosteric pathway of the GroEL reaction cycle, thus trapping an early allosteric intermediate. Addition of GroES is able to overcome this block by binding this intermediate and pulling the allosteric pathway to completion via mass action, explaining how bacterial cells expressing this protein as their only chaperonin are viable.
Translated title of the contributionCharacterisation of a GroEL single-ring mutant that supports growth of Escherichia coli and has GroES-dependent ATPase activity
Original languageEnglish
Pages (from-to)1271 - 1283
Number of pages13
JournalJournal of Molecular Biology
Volume396 (5)
DOIs
Publication statusPublished - Mar 2010

Bibliographical note

Other: First published on-line 16th December 2009

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