title: Molecular characterization of the group II chaperonin from the hyperthermophilic archaeum Pyrococcus horikoshii OT3.
abstract: The group II chaperonin from the hyperthermophilic archaeum Pyrococcus horikoshii OT3 (PhCPN) and its functional cooperation with the cognate prefoldin were investigated. PhCPN existed as a homo-oligomer in a double-ring structure, which protected the citrate synthase of a porcine heart from thermal aggregation at 45 degrees C, and did the same on the isopropylmalate dehydrogenase (IPMDH) of a thermophilic bacterium, Thermus thermophilus HB8, at 90 degrees C. PhCPN also enhanced the refolding of green fluorescent protein (GFP), which had been unfolded by low pH, in an ATP-dependent manner. Unexpectedly, functional cooperation between PhCPN and Pyrococcus prefoldin (PhPFD) in the refolding of GFP was not observed. Instead, cooperation between PhCPN and PhPFD was observed in the refolding of IPMDH unfolded with guanidine hydrochloride. Although PhCPN alone was not effective in the refolding of IPMDH, the refolding efficiency was enhanced by the cooperation of PhCPN with PhPFD.
authors.affiliation: Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan.
authors.forename: Mina
authors.initials: M
authors.lastname: Okochi
authors.forename: Hiroki
authors.initials: H
authors.lastname: Matsuzaki
authors.forename: Tomoko
authors.initials: T
authors.lastname: Nomura
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Ishii
authors.forename: Masafumi
authors.initials: M
authors.lastname: Yohda
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D018833:Chaperonins / Q000737:chemistry* / Q000235:genetics*; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D019791:Guanidine / Q000737:chemistry; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008854:Microscopy, Electron; D018832:Molecular Chaperones / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D017510:Protein Folding; D019711:Pyrococcus / Q000378:metabolism; D043902:Pyrococcus horikoshii / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013552:Swine; D013696:Temperature; D013997:Time Factors
chemical_list: D018832:Molecular Chaperones; C112806:prefoldin; D049452:Green Fluorescent Proteins; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D018833:Chaperonins; D019791:Guanidine