2OP9

Substrate Specificity Profiling and Identification of a New Class of Inhibitor for the Major Protease of the SARS Coronavirus

2OP9 の概要

• エントリーDOI: 10.2210/pdb2op9/pdb
• 分子名称: Replicase polyprotein 1ab (pp1ab, ORF1AB) 3C-like proteinase (3CL-PRO, 3CLp), NALPHA-[(BENZYLOXY)CARBONYL]-N-[(1R)-4-HYDROXY-1-METHYL-2-OXOBUTYL]-L-PHENYLALANINAMIDE (3 entities in total)
• 機能のキーワード: sars, 3clpro, proteinase, protease, hydrolase
• 由来する生物種: SARS coronavirus
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 67627.16

構造登録者

Craik, C.S.,Goetz, D.H. (登録日: 2007-01-27, 公開日: 2007-07-17, 最終更新日: 2024-12-25)

主引用文献

Goetz, D.H.,Choe, Y.,Hansell, E.,Chen, Y.T.,McDowell, M.,Jonsson, C.B.,Roush, W.R.,McKerrow, J.,Craik, C.S.
Substrate Specificity Profiling and Identification of a New Class of Inhibitor for the Major Protease of the SARS Coronavirus.
Biochemistry, 46:8744-8752, 2007

PubMed Abstract: Severe acute respiratory syndrome (SARS) is an emerging infectious disease associated with a high rate of mortality. The SARS-associated coronavirus (SARS-CoV) has been identified as the etiological agent of the disease. Although public health procedures have been effective in combating the spread of SARS, concern remains about the possibility of a recurrence. Various approaches are being pursued for the development of efficacious therapeutics. One promising approach is to develop small molecule inhibitors of the essential major polyprotein processing protease 3Clpro. Here we report a complete description of the tetrapeptide substrate specificity of 3Clpro using fully degenerate peptide libraries consisting of all 160,000 possible naturally occurring tetrapeptides. The substrate specificity data show the expected P1-Gln P2-Leu specificity and elucidate a novel preference for P1-His containing substrates equal to the expected preference for P1-Gln. These data were then used to develop optimal substrates for a high-throughput screen of a 2000 compound small-molecule inhibitor library consisting of known cysteine protease inhibitor scaffolds. We also report the 1.8 A X-ray crystal structure of 3Clpro bound to an irreversible inhibitor. This inhibitor, an alpha,beta-epoxyketone, inhibits 3Clpro with a k3/Ki of 0.002 microM(-1) s(-1) in a mode consistent with the substrate specificity data. Finally, we report the successful rational improvement of this scaffold with second generation inhibitors. These data provide the foundation for a rational small-molecule inhibitor design effort based upon the inhibitor scaffold identified, the crystal structure of the complex, and a more complete understanding of P1-P4 substrate specificity.

PubMed: 17605471
DOI: 10.1021/bi0621415

実験手法

X-RAY DIFFRACTION (1.8 Å)