Abstract
This protocol describes an in vitro fluorogenic assay to measure the proteolytic activity and identify inhibitors of M pro , the main protease produced by SARS-CoV-2 (Severe acute respiratory syndrome coronavirus 2). Studies to identify potential inhibitors of M pro mainly rely on in silico molecular dynamics simulations or on FRET (Fluorescence Resonance Energy Transfer) substrates. The protocol is based on an aminomethyl coumarin substrate. High sensitivity, specificity, and an easily detectable fluorescent read-out are the advantages offered by this rapid assay, which allows high throughput screening of new M pro inhibitors.
Keywords: Biotechnology and bioengineering; Chemistry; Clinical Protocol; Health Sciences; High Throughput Screening; Molecular/Chemical Probes; Protein Biochemistry.
【저자키워드】 Health Sciences, High Throughput Screening, Chemistry, clinical protocol, Protein Biochemistry, Biotechnology and bioengineering, Molecular/Chemical Probes, 【초록키워드】 severe acute respiratory syndrome coronavirus 2, SARS-CoV-2, coronavirus, protocol, coumarin, Biotechnology, molecular dynamics, protease, in vitro, in silico, severe acute respiratory syndrome Coronavirus, inhibitors, Molecular dynamics simulation, sensitivity, specificity, clinical, Chemistry, Fluorescence, inhibitor, bioengineering, energy transfer, energy, acute respiratory syndrome, acute respiratory syndrome coronavirus, acute respiratory syndrome coronavirus 2, M pro, proteolytic activity, fluorescent, throughput, substrates, resonance, produced, identify, detectable, offered, 【제목키워드】 SARS-CoV-2, inhibitor, in vitro activity,