Gentamicin inhibits HSP70-assisted protein folding by interfering with substrate recognition

Yamamoto, Soh; Nakano, Shunsuke; Owari, Kensuke; Fuziwara, Kazuhiko; Ogawa, Nobuaki; Otaka, Michiro; Tamaki, Kumiko; Watanabe, Sumio; Komatsuda, Atsushi; Wakui, Hideki; Sawada, Ken-ichi; Kubota, Hiroshi; Itoh, Hideaki

doi:10.1016/j.febslet.2009.12.021

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Volume 584, Issue 4 , Pages 645-651, 19 February 2010

Gentamicin inhibits HSP70-assisted protein folding by interfering with substrate recognition

Edited by Miguel De la Rosa

Soh Yamamoto
- Department of Life Science, Faculty of Engineering and Resource Science, Akita University, 1-1 Tegata Gakuen Town, Akita City 010-8502, Japan
Shunsuke Nakano
- Department of Life Science, Faculty of Engineering and Resource Science, Akita University, 1-1 Tegata Gakuen Town, Akita City 010-8502, Japan
Kensuke Owari
- Department of Life Science, Faculty of Engineering and Resource Science, Akita University, 1-1 Tegata Gakuen Town, Akita City 010-8502, Japan
Kazuhiko Fuziwara
- Department of Life Science, Faculty of Engineering and Resource Science, Akita University, 1-1 Tegata Gakuen Town, Akita City 010-8502, Japan
Nobuaki Ogawa
- Department of Life Science, Faculty of Engineering and Resource Science, Akita University, 1-1 Tegata Gakuen Town, Akita City 010-8502, Japan
Michiro Otaka
- Department of Gastroenterology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-Ku, Tokyo, Japan
Kumiko Tamaki
- Department of Gastroenterology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-Ku, Tokyo, Japan
Sumio Watanabe
- Department of Gastroenterology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-Ku, Tokyo, Japan
Atsushi Komatsuda
- Third Department of Internal Medicine, Akita University School of Medicine, Akita 010-8543, Japan
Hideki Wakui
- Third Department of Internal Medicine, Akita University School of Medicine, Akita 010-8543, Japan
Ken-ichi Sawada
- Third Department of Internal Medicine, Akita University School of Medicine, Akita 010-8543, Japan
Hiroshi Kubota
- Department of Life Science, Faculty of Engineering and Resource Science, Akita University, 1-1 Tegata Gakuen Town, Akita City 010-8502, Japan
Hideaki Itoh
- Department of Life Science, Faculty of Engineering and Resource Science, Akita University, 1-1 Tegata Gakuen Town, Akita City 010-8502, Japan
- Corresponding author. Fax: +81 18 883 3041.

Received 29 September 2009; received in revised form 24 November 2009; accepted 10 December 2009. published online 22 December 2009.

Abstract

We previously reported that gentamicin (GM) specifically binds to heat-shock protein with subunit molecular masses of 70kDa (HSP70). In the present study, we have investigated the effects of GM binding on HSP70-assisted protein folding in vitro. The C-terminal, and not the N-terminal of HSP70 was found to bind to GM. GM significantly suppressed refolding of firefly luciferase in the presence of HSP70 and HSP40, although the ATPase activity of HSP70 was unaffected by GM. A surface plasmon resonance analysis revealed that GM specifically interferes with the binding of HSP70 to a model peptide that mimics the exposed hydrophobic surface of the folding intermediates. These results indicated that GM inhibits the chaperone activity of HSP70 and may suppress protein folding via inhibition of HSP70 in vivo.