Archaeal group II chaperonin mediates protein folding in the cis-cavity without a detachable GroES-like co-chaperonin

被引:43
作者
Yoshida, T
Kawaguchi, R
Taguchi, H
Yoshida, M
Yasunaga, T
Wakabayashi, T
Yohda, M
Maruyama, T
机构
[1] Marine Biotechnol Inst Co Ltd, Kamaishi Labs, Kamaishi, Iwate 0260001, Japan
[2] Kitasato Univ, Sch Fisheries Sci, Sanriku, Iwate 0220101, Japan
[3] Tokyo Inst Technol, Chem Resources Lab, Midori Ku, Yokohama, Kanagawa 2268503, Japan
[4] Univ Tokyo, Dept Phys, Sch Sci, Bunkyo Ku, Tokyo 1138654, Japan
[5] Tokyo Univ Agr & Technol, Dept Biotechnol & Life Sci, Koganei, Tokyo 1848588, Japan
关键词
archaea; central cavity; cis-folding; group II chaperonin; protein folding mechanism;
D O I
10.1006/jmbi.2001.5220
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Group II chaperonins of archaea and eukaryotes are distinct from group chaperonins of bacteria. Whereas group I chaperonins require the co-chaperonin Cpn-10 or GroES for protein folding, no co-chaperonin has been known for group II. The protein folding mechanism of group II chaperonins is not yet clear. To understand this mechanism, we examined protein refolding by the recombinant alpha or beta-subunit chaperonin homo-oligomer (alpha16mer and beta16mer) from a hyperthermoplilic archaeum, Thermococcus strain KS-1, using a model substrate, green fluorescent protein (GFP). The alpha16mer and beta16mer captured the non-native GFP and promoted its refolding without any co-chaperonin in an ATP dependent manner. A non-hydrolyzable ATP analog, AMP-PNP, induced the GFP refolding mediated by beta16mer but not by the alpha16mer. A mutant alpha-subunit chaperonin homo-oligomer (trap-alpha) could capture the non-native protein but lacked the ability to refold it. Although trap-a suppressed ATP-dependent refolding of GFP mediated by alpha16mer or beta16mer, it did not affect the AMP-PNP-dependent refolding. This indicated that the GFP refolding mediated by beta16mer with AMP-PNP was not accessible to the trap-alpha. Gel filtration chromatography and a protease protection experiment revealed that this refolded GFP, in the presence of AMP-PNP, was associated with beta16mer. After the completion of GFP refolding mediated by beta16mer with AMP-PNP, addition of ATP induced an additional refolding of GFP. Furthermore, the beta16mer preincubated with AMP-PNP showed the ability to capture the non-native GFP. These suggest that AMP-PNP induced one of two chaperonin rings (cis-ring) to close and induced protein refolding in this ring, and that the other ring (trans-ring) could capture the unfolded GFP which was refolded by adding ATP. The present data indicate that, in the group II chaperonin of Thermococcus strain KS-1, the protein folding proceeds in its cis-ring in an ATP-dependent fashion without any co-chaperonin. (C) 2002 Academic Press.
引用
收藏
页码:73 / 85
页数:13
相关论文
共 48 条
[1]   Recurrent paralogy in the evolution of archaeal chaperonins [J].
Archibald, JM ;
Logsdon, JM ;
Doolittle, WF .
CURRENT BIOLOGY, 1999, 9 (18) :1053-1056
[2]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[3]   THE CRYSTAL-STRUCTURE OF THE BACTERIAL CHAPERONIN GROEL AT 2.8-ANGSTROM [J].
BRAIG, K ;
OTWINOWSKI, Z ;
HEGDE, R ;
BOISVERT, DC ;
JOACHIMIAK, A ;
HORWICH, AL ;
SIGLER, PB .
NATURE, 1994, 371 (6498) :578-586
[4]   HARMONIC ANALYSIS OF ELECTRON MICROSCOPE IMAGES WITH ROTATIONAL SYMMETRY [J].
CROWTHER, RA ;
AMOS, LA .
JOURNAL OF MOLECULAR BIOLOGY, 1971, 60 (01) :123-&
[5]   Crystal structure of the thermosome, the archaeal chaperonin and homolog of CCT [J].
Ditzel, L ;
Löwe, J ;
Stock, D ;
Stetter, KO ;
Huber, H ;
Huber, R ;
Steinbacher, S .
CELL, 1998, 93 (01) :125-138
[6]   MOLECULAR CHAPERONES [J].
ELLIS, RJ ;
VANDERVIES, SM .
ANNUAL REVIEW OF BIOCHEMISTRY, 1991, 60 :321-347
[7]   Chaperonin-mediated folding in the eukaryotic cytosol proceeds through rounds of release of native and nonnative forms [J].
Farr, GW ;
Scharl, EC ;
Schumacher, RJ ;
Sondek, S ;
Horwich, AL .
CELL, 1997, 89 (06) :927-937
[8]  
Fenton WA, 1997, PROTEIN SCI, V6, P743
[9]   Principles of chaperone-assisted protein folding: Differences between in vitro and in vivo mechanisms [J].
Frydman, J ;
Hartl, FU .
SCIENCE, 1996, 272 (5267) :1497-1502
[10]   Group II chaperonin in a thermophilic methanogen, Methanococcus thermolithotrophicus -: Chaperone activity and filament-forming ability [J].
Furutani, M ;
Iida, T ;
Yoshida, T ;
Maruyama, T .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (43) :28399-28407