Near-atomic resolution structural model of the yeast 26S proteasome

被引:207
作者
Beck, Florian [1 ]
Unverdorben, Pia [1 ]
Bohn, Stefan [1 ]
Schweitzer, Andreas [1 ]
Pfeifer, Guenter [1 ]
Sakata, Eri [1 ]
Nickell, Stephan [1 ]
Plitzko, Juergen M. [1 ]
Villa, Elizabeth [1 ]
Baumeister, Wolfgang [1 ]
Foerster, Friedrich [1 ]
机构
[1] Max Planck Inst Biochem, Dept Mol Struct Biol, D-82152 Martinsried, Germany
关键词
protein degradation; electron microscopy; deubiquitylating enzyme; REGULATORY PARTICLE; MOLECULAR ARCHITECTURE; CRYSTAL-STRUCTURE; 20S PROTEASOME; MICROSCOPY; DOMAIN; DEGRADATION; RPN11; CORE; DEUBIQUITINATION;
D O I
10.1073/pnas.1213333109
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The 26S proteasome operates at the executive end of the ubiquitin-proteasome pathway. Here, we present a cryo-EM structure of the Saccharomyces cerevisiae 26S proteasome at a resolution of 7.4 angstrom or 6.7 angstrom ( Fourier-Shell Correlation of 0.5 or 0.3, respectively). We used this map in conjunction with molecular dynamics-based flexible fitting to build a near-atomic resolution model of the holocomplex. The quality of the map allowed us to assign alpha-helices, the predominant secondary structure element of the regulatory particle subunits, throughout the entire map. We were able to determine the architecture of the Rpn8/Rpn11 heterodimer, which had hitherto remained elusive. The MPN domain of Rpn11 is positioned directly above the AAA-ATPase N-ring suggesting that Rpn11 deu-biquitylates substrates immediately following commitment and prior to their unfolding by the AAA-ATPase module. The MPN domain of Rpn11 dimerizes with that of Rpn8 and the C-termini of both subunits form long helices, which are integral parts of a coiled-coil module. Together with the C-terminal helices of the six PCI-domain subunits they form a very large coiled-coil bundle, which appears to serve as a flexible anchoring device for all the lid subunits.
引用
收藏
页码:14870 / 14875
页数:6
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