Protein Science, Vol 5, Issue 3 478-487, Copyright © 1996 by Cold Spring Harbor Laboratory Press

ARTICLE

Influence of the GroE molecular chaperone machine on the in vitro refolding of Escherichia coli {beta}-galactosidase

A. AYLING and F. BANEYX
Department of Chemical Engineering, University of Washington, Seattle, Washington 98195-1750

We have studied the effect of the components of the GroE molecular chaperone machine on the refolding of the Escherichia coli enzyme {beta}-galactosidase, a tetrameric protein whose 116-kDa protomers should not completely fit within the central cavity of the GroEL toroid. In the absence of other additives, GroEL formed a weak complex with chemically denatured {beta}-galactosidase, reduced its propensity to aggregate, and increased the recovery yields of active enzyme twofold without altering its folding pathway. When present together with the chaperonin, ATP--and to a lesser extent AMP-PNP--reduced the recovery yields and led to the resumption of aggregation. The use of the complete chaperonin system (GroEL, GroES, and ATP) eliminated the GroEL-mediated increase in recovery and folding proceeded less efficiently than in buffer alone. This unusual behavior can be explained in terms of a chaperonin ``buffering'' effect and the different affinities of GroE complexes for denatured {beta}-galactosidase.
CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				M. M. Carrio and A. Villaverde Localization of Chaperones DnaK and GroEL in Bacterial Inclusion Bodies J. Bacteriol., May 15, 2005; 187(10): 3599 - 3601. [Abstract] [Full Text] [PDF]

				O. V. Polyakova, O. Roitel, R. A. Asryants, A. A. Poliakov, G. Branlant, and V. I. Muronetz Misfolded forms of glyceraldehyde-3-phosphate dehydrogenase interact with GroEL and inhibit chaperonin-assisted folding of the wild-type enzyme Protein Sci., April 1, 2005; 14(4): 921 - 928. [Abstract] [Full Text] [PDF]

				N. Tanaka, Y. Tani, H. Hattori, T. Tada, and S. Kunugi Interaction of the N-terminal domain of Escherichia coli heat-shock protein ClpB and protein aggregates during chaperone activity Protein Sci., December 1, 2004; 13(12): 3214 - 3221. [Abstract] [Full Text] [PDF]

				J. R. Shearstone and F. Baneyx Biochemical Characterization of the Small Heat Shock Protein IbpB from Escherichia coli J. Biol. Chem., April 9, 1999; 274(15): 9937 - 9945. [Abstract] [Full Text] [PDF]

				P. J. Muchowski and J. I. Clark ATP-enhanced molecular chaperone functions of the small heat shock protein human alpha B crystallin PNAS, February 3, 1998; 95(3): 1004 - 1009. [Abstract] [Full Text] [PDF]

				C. Frieden and A. C. Clark Protein folding: How the mechanism of GroEL action is defined by kinetics PNAS, May 27, 1997; 94(11): 5535 - 5538. [Abstract] [Full Text] [PDF]

				J. G. Thomas and F.ço. Baneyx Protein Misfolding and Inclusion Body Formation in Recombinant Escherichia coli Cells Overexpressing Heat-shock Proteins J. Biol. Chem., May 10, 1996; 271(19): 11141 - 11147. [Abstract] [Full Text] [PDF]