Next Article in Journal
Role of mTOR Signaling in Tumor Microenvironment: An Overview
Next Article in Special Issue
Etoposide-Induced Apoptosis in Cancer Cells Can Be Reinforced by an Uncoupled Link between Hsp70 and Caspase-3
Previous Article in Journal
Cellular Pharmacology of Palladinum(III) Hematoporphyrin IX Complexes: Solution Stability, Antineoplastic and Apoptogenic Activity, DNA Binding, and Processing of DNA-Adducts
Previous Article in Special Issue
The Role of the Arginine in the Conserved N-Terminal Domain RLFDQxFG Motif of Human Small Heat Shock Proteins HspB1, HspB4, HspB5, HspB6, and HspB8
Article Menu
Issue 8 (August) cover image

Export Article

Open AccessArticle
Int. J. Mol. Sci. 2018, 19(8), 2452; https://doi.org/10.3390/ijms19082452

Expression, Functional Characterization, and Preliminary Crystallization of the Cochaperone Prefoldin from the Thermophilic Fungus Chaetomium thermophilum

1
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Naka, Koganei, Tokyo 184-8588, Japan
2
Instrumentation Analysis Center, Tokyo University of Agriculture and Technology, Naka, Koganei, Tokyo 184-8588, Japan
3
Department of Molecular Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto 860-0811, Japan.
4
Core Research for Evolutional Science and Technology (CREST), JST, Kawaguchi, Saitama 332-0012, Japan
5
Exploratory Research Center on Life and Living Systems, National Institutes of Natural Sciences, Myodaiji, Okazaki 444-8787, Japan
6
Structural Biology Research Center, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya, Aichi 464-8601, Japan
7
Departamento de Estructura de Macromoléculas, Centro Nacional de Biotecnología (CNB-CSIC), Madrid 28049, Spain
*
Author to whom correspondence should be addressed.
Received: 4 June 2018 / Revised: 15 August 2018 / Accepted: 15 August 2018 / Published: 19 August 2018
(This article belongs to the Special Issue Molecular Chaperones)
Full-Text   |   PDF [2546 KB, uploaded 19 August 2018]   |  

Abstract

Prefoldin is a hexameric molecular chaperone found in the cytosol of archaea and eukaryotes. Its hexameric complex is built from two related classes of subunits, and has the appearance of a jellyfish: Its body consists of a double β-barrel assembly with six long tentacle-like coiled coils protruding from it. Using the tentacles, prefoldin captures an unfolded protein substrate and transfers it to a group II chaperonin. Based on structural information from archaeal prefoldins, mechanisms of substrate recognition and prefoldin-chaperonin cooperation have been investigated. In contrast, the structure and mechanisms of eukaryotic prefoldins remain unknown. In this study, we succeeded in obtaining recombinant prefoldin from a thermophilic fungus, Chaetomium thermophilum (CtPFD). The recombinant CtPFD could not protect citrate synthase from thermal aggregation. However, CtPFD formed a complex with actin from chicken muscle and tubulin from porcine brain, suggesting substrate specificity. We succeeded in observing the complex formation of CtPFD and the group II chaperonin of C. thermophilum (CtCCT) by atomic force microscopy and electron microscopy. These interaction kinetics were analyzed by surface plasmon resonance using Biacore. Finally, we have shown the transfer of actin from CtPFD to CtCCT. The study of the folding pathway formed by CtPFD and CtCCT should provide important information on mechanisms of the eukaryotic prefoldin–chaperonin system. View Full-Text
Keywords: chaperone; chaperonin; Chaetomium thermophilum; proteostasis; interaction; folding chaperone; chaperonin; Chaetomium thermophilum; proteostasis; interaction; folding
Figures

Graphical abstract

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Supplementary material

SciFeed

Share & Cite This Article

MDPI and ACS Style

Morita, K.; Yamamoto, Y.Y.; Hori, A.; Obata, T.; Uno, Y.; Shinohara, K.; Noguchi, K.; Noi, K.; Ogura, T.; Ishii, K.; Kato, K.; Kikumoto, M.; Arranz, R.; Valpuesta, J.M.; Yohda, M. Expression, Functional Characterization, and Preliminary Crystallization of the Cochaperone Prefoldin from the Thermophilic Fungus Chaetomium thermophilum. Int. J. Mol. Sci. 2018, 19, 2452.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top