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Structure paper

タイトル	An allosteric switch regulates ClpP1P2 protease function as established by cryo-EM and methyl-TROSY NMR.
ジャーナル・号・ページ	Proc Natl Acad Sci U S A, Vol. 117, Issue 11, Page 5895-5906, Year 2020
掲載日	2020年3月17日
著者	Siavash Vahidi / Zev A Ripstein / Jordan B Juravsky / Enrico Rennella / Alfred L Goldberg / Anthony K Mittermaier / John L Rubinstein / Lewis E Kay /
PubMed 要旨	The 300-kDa ClpP1P2 protease from collaborates with the AAA+ (ATPases associated with a variety of cellular activities) unfoldases, ClpC1 and ClpX, to degrade substrate proteins. Unlike in other ...The 300-kDa ClpP1P2 protease from collaborates with the AAA+ (ATPases associated with a variety of cellular activities) unfoldases, ClpC1 and ClpX, to degrade substrate proteins. Unlike in other bacteria, all of the components of the Clp system are essential for growth and virulence of mycobacteria, and their inhibitors show promise as antibiotics. MtClpP1P2 is unique in that it contains a pair of distinct ClpP1 and ClpP2 rings and also requires the presence of activator peptides, such as benzoyl-leucyl-leucine (Bz-LL), for function. Understanding the structural basis for this requirement has been elusive but is critical for the rational design and improvement of antituberculosis (anti-TB) therapeutics that target the Clp system. Here, we present a combined biophysical and biochemical study to explore the structure-dynamics-function relationship in MtClpP1P2. Electron cryomicroscopy (cryo-EM) structures of apo and acyldepsipeptide-bound MtClpP1P2 explain their lack of activity by showing loss of a key β-sheet in a sequence known as the handle region that is critical for the proper formation of the catalytic triad. Methyl transverse relaxation-optimized spectroscopy (TROSY)-based NMR, cryo-EM, and biochemical assays show that, on binding Bz-LL or covalent inhibitors, MtClpP1P2 undergoes a conformational change from an inactive compact state to an active extended structure that can be explained by a modified Monod-Wyman-Changeux model. Our study establishes a critical role for the handle region as an on/off switch for function and shows extensive allosteric interactions involving both intra- and interring communication that regulate MtClpP1P2 activity and that can potentially be exploited by small molecules to target .
リンク	Proc Natl Acad Sci U S A / PubMed:32123115 / PubMed Central
手法	EM (単粒子)
解像度	3.1 - 3.5 Å
構造データ	21197 6vgk EMDB-21197, PDB-6vgk: ClpP1P2 complex from M. tuberculosis 手法: EM (単粒子) / 解像度: 3.1 Å 21198 6vgn EMDB-21198, PDB-6vgn: ClpP1P2 complex from M. tuberculosis bound to ADEP 手法: EM (単粒子) / 解像度: 3.1 Å 21199 6vgq EMDB-21199, PDB-6vgq: ClpP1P2 complex from M. tuberculosis with GLF-CMK bound to ClpP1 手法: EM (単粒子) / 解像度: 3.5 Å
由来	mycobacterium tuberculosis (結核菌) synthetic construct (人工物)
キーワード	HYDROLASE / Complex / protease / ClpP / tuberculosis / HYDROLASE/ANTIBIOTIC / HYDROLASE-ANTIBIOTIC complex