Split conformation of Chaetomium thermophilum Hsp104 disaggregase

Journal Structure 29:1–10 (2021)
Title Split conformation of Chaetomium thermophilum Hsp104 disaggregase
Laboratory JEOL YOKOGUSHI Research Alliance Laboratories〈SA Prof. NAMBA Keiichi〉

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.


Yosuke Inoue (1), Yuya Hanazono (2), Kentaro Noi (3), Akihiro Kawamoto (4), Masato Kimatsuka (5), Ryuhei Harada (5), Kazuki Takeda (2), Ryoichi Kita (1), Natsuki Iwamasa (1), Kyoka Shibata (1), Keiichi Noguchi (1), Yasuteru Shigeta (5), Keiichi Namba (4, 6, 7), Teru Ogura (3), Kunio Miki (2), Kyosuke Shinohara (1), Masafumi Yohda (1)

  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. RIKEN Center for Biosystems Dynamics Research and SPring-8 Center, Suita, Osaka 565-0871, Japan
  7. JEOL YOKOGUSHI Research Alliance Laboratories, Osaka University, Suita, Osaka 565-0871 Japan.
PubMed 33651974