Flavonoid-mediated Inhibition of SARS Coronavirus 3C-like Protease Expressed in Pichia Pastoris

Overview

Journal Biotechnol Lett

Specialties Biochemistry
Biotechnology

Date 2012 Feb 22

PMID 22350287

Citations 164

Authors

Thi Thanh Hanh Nguyen

Hye-Jin Woo

Hee-Kyoung Kang

Van Dao Nguyen

Young-Min Kim

Do-Won Kim

Sul-Ah Ahn

Yongmei Xia

Doman Kim

Affiliations

Soon will be listed here.

Abstract

The 3C-like protease (3CL(pro)) of severe acute respiratory syndrome associated coronavirus (SARS-CoV) is vital for SARS-CoV replication and is a promising drug target. Recombinant 3CL(pro) was expressed in Pichia pastoris GS115 as a 42 kDa protein that displayed a K ( m ) of 15 ± 2 μM with Dabcyl-KTSAVLQSGFRKME-Edans as substrate. Purified 3CL(pro) was used for inhibition and kinetic assays with seven flavonoid compounds. The IC(50) of six flavonoid compounds were 47-381 μM. Quercetin, epigallocatechin gallate and gallocatechin gallate (GCG) displayed good inhibition toward 3CL(pro) with IC(50) values of 73, 73 and 47 μM, respectively. GCG showed a competitive inhibition pattern with K ( i ) value of 25 ± 1.7 μM. In molecular docking experiments, GCG displayed a binding energy of -14 kcal mol(-1) to the active site of 3CL(pro) and the galloyl moiety at 3-OH position was required for 3CL(pro) inhibition activity.

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