[5-TAMRA]/[Lys(BHQ-2)]-CoV Main Protease (Mpro) Substrate – Fluorescent Peptides

Fluorescently labelled substrate for the severe acute respiratory syndrome coronavirus main protease (SARS-CoV Mpro). The substrate sequence is derived from residues P4-P5′ of the SARS-CoV Mpro N-terminal autoprocessing site which has the sequence AVLQSGFRK. SARS-CoV Mpro is a key antiviral target. This peptide contains an N-terminal a 5-carboxytetramethylrhodamine (5-TAMRA), a widely used fluorescent dye which excites at 546 nm and emits at 579 nm and a black hole quencher 2 (BHQ-2) group. This Mpro substrate is used in Mpro inhibition assays. The compound being tested for its inhibitory capacity of Mpro is added alongside [5-TAMRA]/[Lys(BHQ-2)]-CoV, after a set time period, the amount of fluorescence released is read as a percentage inhibition by a plate reader. The fluorescence from 5-TAMRA is efficiently quenched by resonance energy transfer to the BHQ-2 group when the peptide is intact, however upon cleavage of the peptide by Mpro, 5-TAMRA and BHQ-2 are separated, allowing fluorescence to be detected. This therefore represents a useful tool for investigating Mpro activity.

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