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  Efficient Catalysis of Protein Folding by GroEL/ES of the Obligate Chaperonin Substrate MetF

Singh, A. K., Balchin, D., Imamoglu, R., Hayer-Hartl, M., & Hartl, F. U. (2020). Efficient Catalysis of Protein Folding by GroEL/ES of the Obligate Chaperonin Substrate MetF. JOURNAL OF MOLECULAR BIOLOGY, 432(7), 2304-2318. doi:10.1016/j.jmb.2020.02.031.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0006-7F5C-6 Version Permalink: http://hdl.handle.net/21.11116/0000-0006-7F5D-5
Genre: Journal Article

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 Creators:
Singh, Amit K.1, Author              
Balchin, David1, Author              
Imamoglu, Rahmi1, Author              
Hayer-Hartl, Manajit1, Author              
Hartl, F. Ulrich1, Author              
Affiliations:
1Hartl, Franz-Ulrich / Cellular Biochemistry, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565152              

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Free keywords: CRYSTAL-STRUCTURE; METHYLENETETRAHYDROFOLATE REDUCTASE; STRUCTURAL FEATURES; NANO-CAGE; COLI; MECHANISM; CONFORMATION; CONFINEMENT; EXCHANGE; SPECTROSCOPYChaperonins; GroEL; MetF; Single molecule spectroscopy; Hydrogen/deuterium exchange;
 Abstract: The cylindrical chaperonin GroEL and its cofactor GroES mediate ATP-dependent protein folding in Escherichia colt by transiently encapsulating non-native substrate in a nano-cage formed by the GroEL ring cavity and the lid-shaped GroES. Mechanistic studies of GroEL/ES with heterologous protein substrates suggested that the chaperonin is inefficient, typically requiring multiple ATP-dependent encapsulation cycles with only a few percent of protein folded per cycle. Here we analyzed the spontaneous and chaperoninassisted folding of the essential enzyme 5,10-methylenetetrahydrofolate reductase (MetF) of E. coli, an obligate GroEL/ES substrate. We found that MetF, a homotetramer of 33-kDa subunits with (beta/alpha)(8) TIM-barrel fold, populates a kinetically trapped folding intermediate(s) (MetF-I) upon dilution from denaturant that fails to convert to the native state, even in the absence of aggregation. GroEUES recognizes MetF-I and catalyzes rapid folding, with 50% of protein folded in a single round of encapsulation. Analysis by hydrogen/deuterium exchange at peptide resolution showed that the MetF subunit folds to completion in the GroEL/ES nano-cage and binds its cofactor flavin adenine dinucleotide. Rapid folding required the net negative charge character of the wall of the chaperonin cavity. These findings reveal a remarkable capacity of GroEL/ES to catalyze folding of an endogenous substrate protein that would have coevolved with the chaperonin system. (C) 2020 Elsevier Ltd. All rights reserved.

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Language(s): eng - English
 Dates: 2020
 Publication Status: Published in print
 Pages: 15
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: ISI: 000532698400028
DOI: 10.1016/j.jmb.2020.02.031
 Degree: -

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Title: JOURNAL OF MOLECULAR BIOLOGY
Source Genre: Journal
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Publ. Info: 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND : ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
Pages: - Volume / Issue: 432 (7) Sequence Number: - Start / End Page: 2304 - 2318 Identifier: ISSN: 0022-2836