title: Single-molecule analyses of the dynamics of heat shock protein 104 (Hsp104) and protein aggregates.
abstract: Hsp104 solubilizes protein aggregates in cooperation with Hsp70/40. Although the framework of the disaggregase function has been elucidated, the actual process of aggregate solubilization by Hsp104-Hsp70/40 remains poorly understood. Here we developed several methods to investigate the functions of Hsp104 and Hsp70/40 from Saccharomyces cerevisiae, at single-molecule levels. The single-molecule methods, which provide the size distribution of the aggregates, revealed that Hsp70/40 prevented the formation of large aggregates from small aggregates and that the solubilization of the small aggregates required both Hsp104 and Hsp70/40. We directly visualized the individual association-dissociation dynamics of Hsp104 on immobilized aggregates and found that the lifetimes of the Hsp104-aggregate complex are divided into two groups: short (∼4 s) and long (∼30 s). Hsp70/40 stimulated the association of Hsp104 with aggregates and increased the duration of this association. The single-molecule data provide novel insights into the functional mechanism of the Hsp104 disaggregation machine.
authors: From the Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, B-56, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.
authors: Momoko
authors: M
authors: Okuda
authors: From the Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, B-56, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.
authors: Tatsuya
authors: T
authors: Niwa
authors: From the Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, B-56, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan taguchi@bio.titech.ac.jp.
authors: Hideki
authors: H
authors: Taguchi
mesh_terms: D006360:Heat-Shock Proteins / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*
chemical_list: D006360:Heat-Shock Proteins; D029701:Saccharomyces cerevisiae Proteins; C070972:HsP104 protein, S cerevisiae; D008156:Luciferases
keywords: ATPases Associated with Diverse Cellular Activities (AAA); Chaperone; Fluorescence Correlation Spectroscopy (FCS); Hsp104; Protein Aggregation; Single-molecule Biophysics; Total Internal Reflection Fluorescence (TIRF) Microscopy
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