Expression, functional characterization, and preliminary crystallization of the cochaperone prefoldin from the thermophilic fungus chaetomium thermophilum

Autor: Ayaka Hori, Keiichi Noguchi, Yuko Uno, Kentaro Ishii, Mahito Kikumoto, Kento Morita, Teru Ogura, Kentaro Noi, Tomohiro Obata, Rocío Arranz, José M. Valpuesta, Masafumi Yohda, Yohei Y. Yamamoto, Kyosuke Shinohara, Koichi Kato
Přispěvatelé: Ministry of Education, Culture, Sports, Science and Technology (Japan), National Institutes of Natural Sciences (Japan), Japan Science and Technology Agency, Kumamoto University, Ministerio de Economía y Competitividad (España), European Commission
Rok vydání: 2018
Předmět:
Models
Molecular
0301 basic medicine
folding
Protein Folding
Swine
Gene Expression
Chaperone
Chaetomium
environment and public health
Substrate Specificity
Chaperonin
law.invention
lcsh:Chemistry
Chaetomium thermophilum
law
chaperone
Cloning
Molecular
lcsh:QH301-705.5
Spectroscopy
biology
Chemistry
General Medicine
Recombinant Proteins
Computer Science Applications
Recombinant DNA
Crystallization
Protein Binding
chaperonin
Interaction
interaction
macromolecular substances
Article
Catalysis
Fungal Proteins
Inorganic Chemistry
Protein Aggregates
03 medical and health sciences
Animals
Physical and Theoretical Chemistry
Molecular Biology
Actin
proteostasis
Thermophile
Organic Chemistry
Folding
Prefoldin
030104 developmental biology
Tubulin
lcsh:Biology (General)
lcsh:QD1-999
Chaperone (protein)
biology.protein
Biophysics
health occupations
Proteostasis
bacteria
Chickens
Molecular Chaperones
Zdroj: Digital.CSIC. Repositorio Institucional del CSIC
instname
International Journal of Molecular Sciences, Vol 19, Iss 8, p 2452 (2018)
International Journal of Molecular Sciences
Volume 19
Issue 8
Popis: 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.
This research was supported by grants-in-aids for scientific research (JP18H04690, JP16H04572, JP16H00753 and JP15J08261) from the Ministry of Education, Science, Sports, and Culture of Japan, the Joint Studies Program (2017-2018) in the Okazaki BIO-NEXT project of National Institutes of Natural Sciences and a grant from CREST (JPMJCR13M1), the program of the Joint Usage/Research Center for Developmental Medicine (IMEG, Kumamoto University). This work was also supported by the grant BFU2016-75984 (AEI/FEDER, UE).
Databáze: OpenAIRE
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