Probing the dynamic process of encapsulation in Escherichia coli GroEL

Toshifumi Mizuta; Kasumi Ando; Tatsuya Uemura; Yasushi Kawata; Tomohiro Mizobata

doi:10.1371/journal.pone.0078135

Probing the dynamic process of encapsulation in Escherichia coli GroEL

PLoS One. 2013 Oct 30;8(10):e78135. doi: 10.1371/journal.pone.0078135. eCollection 2013.

Authors

Toshifumi Mizuta¹, Kasumi Ando, Tatsuya Uemura, Yasushi Kawata, Tomohiro Mizobata

Affiliation

¹ Department of Biotechnology, Graduate School of Engineering, Tottori, Japan.

Abstract

Kinetic analyses of GroE-assisted folding provide a dynamic sequence of molecular events that underlie chaperonin function. We used stopped-flow analysis of various fluorescent GroEL mutants to obtain details regarding the sequence of events that transpire immediately after ATP binding to GroEL and GroEL with prebound unfolded proteins. Characterization of GroEL CP86, a circularly permuted GroEL with the polypeptide ends relocated to the vicinity of the ATP binding site, showed that GroES binding and protection of unfolded protein from solution is achieved surprisingly early in the functional cycle, and in spite of greatly reduced apical domain movement. Analysis of fluorescent GroEL SR-1 and GroEL D398A variants suggested that among other factors, the presence of two GroEL rings and a specific conformational rearrangement of Helix M in GroEL contribute significantly to the rapid release of unfolded protein from the GroEL apical domain.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Escherichia coli / metabolism*
Escherichia coli Proteins / chemistry*
Escherichia coli Proteins / metabolism*
Heat-Shock Proteins / chemistry*
Heat-Shock Proteins / metabolism*
Kinetics
Protein Structure, Secondary

Substances

Escherichia coli Proteins
GroE protein, E coli
Heat-Shock Proteins

Grants and funding

This study was partially supported by a Grant-in-Aid for Scientific Research (C) (No. 22570119 to TM) from the Japan Society for the Promotion of Science (JSPS) of Japan, a Grant-in-Aid for Scientific Research on Priority Areas (No. 22020025 to YK) from The Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan, and also by grants to TM from the Faculty of Engineering, Tottori University and the Venture Business Laboratory,Tottori University. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.