Design and Synthesis of a Novel Fluorescent Peptide Substrate of SARS-CoⅤ 3CLpro and Studies on Its Enzymatic Kinetics

Chem. J. Chinese Universities

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Design and Synthesis of a Novel Fluorescent Peptide Substrate of SARS-CoⅤ 3CL^pro and Studies on Its Enzymatic Kinetics

WAN Hui-Xin, CHEN Li-Li, LI Xin, WANG Xin, HU Ding-Yu, SHEN Xu, SHEN Jing-Kang

State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Shanghai, 201203, China

Received:2006-02-10 Revised:1900-01-01 Online:2007-01-10 Published:2007-01-10
Contact: SHEN Jing-Kang

Abstract

Abstract: A novel fluorescent peptide substrate, H₂N-E(Eddnp) STLQSGLK(Abz)-CONH₂, based on fluorescence resonance energy transfer(FRET) technique, was designed and synthesized by solid-phase peptide synthesis. Abz/Eddnp was used as the fluorescence donor and quenching pair. The highly fluorescent Abz group was efficiently quenched by energy transfer to the Eddnp group. The fluorescent substrate was specifically hydrolyzed by SARS-CoⅤ 3CL^pro between QS with 15-fold increase in fluorescence and the cleavage site was determined by HPLC-MS. The studies on its enzymatic kinetics(K_m= 525.5 μmol/L and K_cat=1.29 min^-1) elucidate that the synthetic fluorescent peptide substrate is suited to be used as a fluorescent probe in determination of the activity of SARS-CoⅤ 3CL^pro and screening assays for its inhibitors.

Key words: Fluorescence resonance energy transfer(FRET), SARS-CoⅤ 3CL^pro, Fluorescent probe, LC-MS

CLC Number:

O629.72
O643

TrendMD:

WAN Hui-Xin, CHEN Li-Li, LI Xin, WANG Xin, HU Ding-Yu, SHEN Xu, SHEN Jing-Kang. Design and Synthesis of a Novel Fluorescent Peptide Substrate of SARS-CoⅤ 3CL^pro and Studies on Its Enzymatic Kinetics[J]. Chem. J. Chinese Universities, doi: .

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Design and Synthesis of a Novel Fluorescent Peptide Substrate of SARS-CoⅤ 3CL^pro and Studies on Its Enzymatic Kinetics

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