+1 Recommend
0 collections
      • Record: found
      • Abstract: found
      • Article: not found

      Characterization of SARS-CoV main protease and identification of biologically active small molecule inhibitors using a continuous fluorescence-based assay

      , , , , , ,
      Febs Letters
      Federation of European Biochemical Societies. Published by Elsevier B.V.
      SARS-CoV, severe acute respiratory syndrome associated coronavirus, Mpro, main protease, HPLC, high performance liquid chromatography, HTS, high throughput screening, RT-PCR, reverse transcription polymerase chain reaction, PCR, polymerase chain reaction, IPTG, isopropyl β-d-thiogalactoside, DABCYL, 4-(4-dimethylaminophenylazol)benzoyl, EDANS, 5-(2-aminoethylamino)-1-naphthalenesulphonic acid, CPE, cytopathic effect, PRA, plaque reduction assay, EMEM, Eagle's minimal essential medium, FBS, fetal bovine serum, PFU, plaque forming unit, MTT, 3-[4,5-dimethylth-iazol-2-yl]-2,5-diphenyltetrazolium bromide, CMV, cytomegalovirus, Severe acute respiratory syndrome, Coronavirus, 3C-like protease, Main protease, Fluorogenic substrate, Small molecule inhibitor

      Read this article at

          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.


          Severe acute respiratory syndrome associated coronavirus main protease (SARS-CoV M pro) has been proposed as a prime target for anti-SARS drug development. We have cloned and overexpressed the SARS-CoV M pro in Escherichia coli, and purified the recombinant M pro to homogeneity. The kinetic parameters of the recombinant SARS-CoV M pro were characterized by high performance liquid chromatography-based assay and continuous fluorescence-based assay. Two novel small molecule inhibitors of the SARS-CoV M pro were identified by high-throughput screening using an internally quenched fluorogenic substrate. The identified inhibitors have K i values at low μM range with comparable anti-SARS-CoV activity in cell-based assays.

          Related collections

          Author and article information

          FEBS Lett
          FEBS Lett
          Febs Letters
          Federation of European Biochemical Societies. Published by Elsevier B.V.
          25 September 2004
          22 October 2004
          25 September 2004
          : 576
          : 3
          : 325-330
          Department of Microbiology, The University of Hong Kong, Pokfulam, Hong Kong, China
          Author notes
          []Corresponding author. Fax: +852-28167415 rytkao@ 123456hkucc.hku.hk
          Copyright © 2004 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

          Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

          : 22 June 2004
          : 9 September 2004
          : 13 September 2004

          Molecular biology
          sars-cov, severe acute respiratory syndrome associated coronavirus,mpro, main protease,hplc, high performance liquid chromatography,hts, high throughput screening,rt-pcr, reverse transcription polymerase chain reaction,pcr, polymerase chain reaction,iptg, isopropyl β-d-thiogalactoside,dabcyl, 4-(4-dimethylaminophenylazol)benzoyl,edans, 5-(2-aminoethylamino)-1-naphthalenesulphonic acid,cpe, cytopathic effect,pra, plaque reduction assay,emem, eagle's minimal essential medium,fbs, fetal bovine serum,pfu, plaque forming unit,mtt, 3-[4,5-dimethylth-iazol-2-yl]-2,5-diphenyltetrazolium bromide,cmv, cytomegalovirus,severe acute respiratory syndrome,coronavirus,3c-like protease,main protease,fluorogenic substrate,small molecule inhibitor


          Comment on this article