title: Asp-52 in combination with Asp-398 plays a critical role in ATP hydrolysis of chaperonin GroEL.
abstract: The Escherichia coli chaperonin GroEL is a double-ring chaperone that assists protein folding with the aid of GroES and ATP. Asp-398 in GroEL is known as one of the critical residues on ATP hydrolysis because GroEL(D398A) mutant is deficient in ATP hydrolysis (<2% of the wild type) but not in ATP binding. In the archaeal Group II chaperonin, another aspartate residue, Asp-52 in the corresponding E. coli GroEL, in addition to Asp-398 is also important for ATP hydrolysis. We investigated the role of Asp-52 in GroEL and found that ATPase activity of GroEL(D52A) and GroEL(D52A/D398A) mutants was ∼ 20% and <0.01% of wild-type GroEL, respectively, indicating that Asp-52 in E. coli GroEL is also involved in the ATP hydrolysis. GroEL(D52A/D398A) formed a symmetric football-shaped GroEL-GroES complex in the presence of ATP, again confirming the importance of the symmetric complex during the GroEL ATPase cycle. Notably, the symmetric complex of GroEL(D52A/D398A) was extremely stable, with a half-time of ∼ 150 h (∼ 6 days), providing a good model to characterize the football-shaped complex.
authors.affiliation: From the Department of Applied Bioscience, Kanagawa Institute of Technology, 1030 Shimo-ogino, Atsugi, Kanagawa 243-0292, Japan.
authors.forename: Ayumi
authors.initials: A
authors.lastname: Koike-Takeshita
authors.affiliation: Technology Research Association for Next Generation Natural Products Chemistry, AIST Tokyo Waterfront, 2-3-26, Aomi, Koto-ku, Tokyo 135-0064, Japan, and.
authors.forename: Kaoru
authors.initials: K
authors.lastname: Mitsuoka
authors.affiliation: 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.forename: Hideki
authors.initials: H
authors.lastname: Taguchi
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism*; D001224:Aspartic Acid / Q000378:metabolism*; D001665:Binding Sites; D018835:Chaperonin 10 / Q000378:metabolism / Q000648:ultrastructure; D018834:Chaperonin 60 / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D004926:Escherichia coli / Q000378:metabolism*; D006868:Hydrolysis; D008291:Malate Dehydrogenase / Q000378:metabolism; D050505:Mutant Proteins / Q000378:metabolism; D016624:Negative Staining; D017510:Protein Folding; D055550:Protein Stability; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship; D013884:Thiosulfate Sulfurtransferase / Q000378:metabolism
chemical_list: D018835:Chaperonin 10; D018834:Chaperonin 60; D050505:Mutant Proteins; D021122:Protein Subunits; D001224:Aspartic Acid; D000255:Adenosine Triphosphate; D008291:Malate Dehydrogenase; D013884:Thiosulfate Sulfurtransferase
keywords: ATPase; Chaperone; Chaperonin; GroEL; GroES; Protein Aggregation; Protein Folding; Protein Misfolding