Split conformation of Chaetomium thermophilum Hsp104 disaggregase

Yosuke Inoue; Yuya Hanazono; Kentaro Noi; Akihiro Kawamoto; Masato Kimatsuka; Ryuhei Harada; Kazuki Takeda; Ryoichi Kita; Natsuki Iwamasa; Kyoka Shibata; Keiichi Noguchi; Yasuteru Shigeta; Keiichi Namba; Teru Ogura; Kunio Miki; Kyosuke Shinohara; Masafumi Yohda

doi:10.1016/j.str.2021.02.002

Split conformation of Chaetomium thermophilum Hsp104 disaggregase

Structure. 2021 Jul 1;29(7):721-730.e6. doi: 10.1016/j.str.2021.02.002. Epub 2021 Mar 1.

Authors

Yosuke Inoue¹, Yuya Hanazono², Kentaro Noi³, Akihiro Kawamoto⁴, Masato Kimatsuka⁵, Ryuhei Harada⁵, Kazuki Takeda², Ryoichi Kita¹, Natsuki Iwamasa¹, Kyoka Shibata¹, Keiichi Noguchi¹, Yasuteru Shigeta⁵, Keiichi Namba⁶, Teru Ogura³, Kunio Miki², Kyosuke Shinohara⁷, Masafumi Yohda¹

Affiliations

¹ Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan.
² Department of Chemistry, Graduate School of Science, Kyoto University, Kyoto, Kyoto 606-8502, Japan.
³ Department of Molecular Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Kumamoto 860-0811, Japan.
⁴ Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
⁵ Division of Life Science, Center for Computational Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8577, Japan.
⁶ Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan; RIKEN Center for Biosystems Dynamics Research and SPring-8 Center, Suita, Osaka 565-0871, Japan; JEOL YOKOGUSHI Research Alliance Laboratories, Osaka University, Suita, Osaka 565-0871 Japan.
⁷ Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Koganei, Tokyo 184-8588, Japan. Electronic address: k_shino@cc.tuat.ac.jp.

PMID: 33651974
DOI: 10.1016/j.str.2021.02.002

Abstract

Hsp104 and its bacterial homolog ClpB form hexameric ring structures and mediate protein disaggregation. The disaggregated polypeptide is thought to thread through the central channel of the ring. However, the dynamic behavior of Hsp104 during disaggregation remains unclear. Here, we reported the stochastic conformational dynamics and a split conformation of Hsp104 disaggregase from Chaetomium thermophilum (CtHsp104) in the presence of ADP by X-ray crystallography, cryo-electron microscopy (EM), and high-speed atomic force microscopy (AFM). ADP-bound CtHsp104 assembles into a 6₅ left-handed spiral filament in the crystal structure at a resolution of 2.7 Å. The unit of the filament is a hexamer of the split spiral structure. In the cryo-EM images, staggered and split hexameric rings were observed. Further, high-speed AFM observations showed that a substrate addition enhanced the conformational change and increased the split structure's frequency. Our data suggest that split conformation is an off-pathway state of CtHsp104 during disaggregation.

Keywords: Hsp104; molecular chaperone.

Publication types

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

MeSH terms

Adenosine Diphosphate / metabolism*
Chaetomium / chemistry
Chaetomium / metabolism*
Cryoelectron Microscopy
Crystallography, X-Ray
Fungal Proteins / chemistry
HSP40 Heat-Shock Proteins / chemistry*
HSP40 Heat-Shock Proteins / metabolism*
Microscopy, Atomic Force
Models, Molecular
Protein Aggregates
Protein Binding
Protein Conformation
Protein Domains
Protein Multimerization

Substances

Fungal Proteins
HSP40 Heat-Shock Proteins
Protein Aggregates
Adenosine Diphosphate

Supplementary concepts

Chaetomium thermophilum