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Title: Effects of C-terminal Truncation of Chaperonin GroEL on the Yield of In-cage Folding of the Green Fluorescent Protein.
Authors: Ishino, So
Kawata, Yasushi
Taguchi, Hideki
Kajimura, Naoko
Matsuzaki, Katsumi  kyouindb  KAKEN_id  orcid (unconfirmed)
Hoshino, Masaru  kyouindb  KAKEN_id  orcid (unconfirmed)
Author's alias: 星野, 大
Keywords: ATP
Escherichia coli (E. coli)
molecular chaperone
protein folding
Issue Date: 12-Jun-2015
Publisher: American Society for Biochemistry and Molecular Biology
Journal title: The Journal of biological chemistry
Volume: 290
Issue: 24
Start page: 15042
End page: 15051
Abstract: Chaperonin GroEL from Escherichia coli consists of two heptameric rings stacked back-to-back to form a cagelike structure. It assists in the folding of substrate proteins in concert with the co-chaperonin GroES by incorporating them into its large cavity. The mechanism underlying the incorporation of substrate proteins currently remains unclear. The flexible C-terminal residues of GroEL, which are invisible in the x-ray crystal structure, have recently been suggested to play a key role in the efficient encapsulation of substrates. These C-terminal regions have also been suggested to separate the double rings of GroEL at the bottom of the cavity. To elucidate the role of the C-terminal regions of GroEL on the efficient encapsulation of substrate proteins, we herein investigated the effects of C-terminal truncation on GroE-mediated folding using the green fluorescent protein (GFP) as a substrate. We demonstrated that the yield of in-cage folding mediated by a single ring GroEL (SR1) was markedly decreased by truncation, whereas that mediated by a double ring football-shaped complex was not affected. These results suggest that the C-terminal region of GroEL functions as a barrier between rings, preventing the leakage of GFP through the bottom space of the cage. We also found that once GFP folded into its native conformation within the cavity of SR1 it never escaped even in the absence of the C-terminal tails. This suggests that GFP molecules escaped through the pore only when they adopted a denatured conformation. Therefore, the folding and escape of GFP from C-terminally truncated SR1·GroES appeared to be competing with each other.
Rights: This research was originally published in [The Journal of Biological Chemistry, 290, 15042-15051. doi: 10.1074/jbc.M114.633636.] © the American Society for Biochemistry and Molecular Biology
This is not the published version. Please cite only the published version.
DOI(Published Version): 10.1074/jbc.M114.633636
PubMed ID: 25887400
Appears in Collections:Journal Articles

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