Peptide aldehyde inhibitors challenge the substrate specificity of the SARS-coronavirus main protease

doi:10.1016/j.antiviral.2011.08.001

	Peptide aldehyde inhibitors challenge the substrate specificity of the SARS-coronavirus main protease
	Zhu, Lili 2; George, Shyla 2; Schmidt, Marco F.3; Al-Gharabli, Samer I.4; Rademann, Joerg 1,3; Hilgenfeld, Rolf 2,5,6
刊名	ANTIVIRAL RESEARCH
	2011-11
卷号	92 期号:2 页码:204-212
关键词	Aldehyde inhibitor Antiviral drug design Cysteine protease Methionine-aspartic acid interaction X-ray crystallography
ISSN号	0166-3542
DOI	10.1016/j.antiviral.2011.08.001
文献子类	Article
英文摘要	SARS coronavirus main protease (SARS-CoV M(pro)) is essential for the replication of the virus and regarded as a major antiviral drug target. The enzyme is a cysteine protease, with a catalytic dyad (Cys-145/His-41) in the active site. Aldehyde inhibitors can bind reversibly to the active-site sulfhydryl of SARS-CoV M(pro). Previous studies using peptidic substrates and inhibitors showed that the substrate specificity of SARS-CoV M(pro) requires glutamine in the P1 position and a large hydrophobic residue in the P2 position. We determined four crystal structures of SARS-CoV M(pro) in complex with pentapeptide aldehydes (Ac-EST-LQ-H, Ac-NSFSQ-H, Ac-DSFDQ-H, and Ac-NSTSQ-H). Kinetic data showed that all of these aldehydes exhibit inhibitory activity towards SARS-CoV M(pro), with K(i) values in the mu M range. Surprisingly, the X-ray structures revealed that the hydrophobic S2 pocket of the enzyme can accommodate serine and even aspartic-acid side-chains in the P2 positions of the inhibitors. Consequently, we reassessed the substrate specificity of the enzyme by testing the cleavage of 20 different tetradecapeptide substrates with varying amino-acid residues in the P2 position. The cleavage efficiency for the substrate with serine in the P2 position was 160-times lower than that for the original substrate (P2 = Leu); furthermore, the substrate with aspartic acid in the P2 position was not cleaved at all. We also determined a crystal structure of SARS-CoV M(pro) in complex with aldehyde Cm-FF-H, which has its P1-phenylalanine residue bound to the relatively hydrophilic S1 pocket of the enzyme and yet exhibits a high inhibitory activity against SARS-CoV M(pro), with K(i) = 2.24 +/- 0.58 mu M. These results show that the stringent substrate specificity of the SARS-CoV M(pro) with respect to the P1 and P2 positions can be overruled by the highly electrophilic character of the aldehyde warhead, thereby constituting a deviation from the dogma that peptidic inhibitors need to correspond to the observed cleavage specificity of the target protease. (C) 2011 Elsevier B.V. All rights reserved.
资助项目	Deutsche Forschungsgemeinschaft[Hi611/4] ; Deutsche Forschungsgemeinschaft[Ra895/2] ; Sino-German Center for the Promotion of Research, Beijing[00000000] ; European Commission[SP22-CT-2004-003831] ; European Commission[HEALTH-F3-2010-260644] ; Chinese Academy of Sciences[2010T1S6]
WOS关键词	COV M-PRO ; 3C-LIKE PROTEINASE ; CRYSTAL-STRUCTURES ; DESIGN ; DISCOVERY ; COMPLEX ; ESTERS ; MODEL ; DIMER ; VIEW
WOS研究方向	Pharmacology & Pharmacy ; Virology
语种	英语
出版者	ELSEVIER SCIENCE BV
WOS记录号	WOS:000297427200008
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/278347]
专题	中国科学院上海药物研究所
通讯作者	Hilgenfeld, Rolf
作者单位	1.Univ Leipzig, Inst Pharm, D-04103 Leipzig, Germany; 2.Med Univ Lubeck, Inst Biochem, Ctr Struct & Cell Biol Med, D-23538 Lubeck, Germany; 3.Leibniz Inst Mol Pharmacol FMP, D-13125 Berlin, Germany; 4.German Jordanian Univ, Chem Pharmaceut Engn Dept, Amman 11180, Jordan; 5.Chinese Acad Sci, Shanghai Inst Mat Med, Shanghai 201203, Peoples R China; 6.DESY, Lab Struct Biol Infect & Inflammat, D-22603 Hamburg, Germany
推荐引用方式 GB/T 7714	Zhu, Lili,George, Shyla,Schmidt, Marco F.,et al. Peptide aldehyde inhibitors challenge the substrate specificity of the SARS-coronavirus main protease[J]. ANTIVIRAL RESEARCH,2011,92(2):204-212.
APA	Zhu, Lili,George, Shyla,Schmidt, Marco F.,Al-Gharabli, Samer I.,Rademann, Joerg,&Hilgenfeld, Rolf.(2011).Peptide aldehyde inhibitors challenge the substrate specificity of the SARS-coronavirus main protease.ANTIVIRAL RESEARCH,92(2),204-212.
MLA	Zhu, Lili,et al."Peptide aldehyde inhibitors challenge the substrate specificity of the SARS-coronavirus main protease".ANTIVIRAL RESEARCH 92.2(2011):204-212.