RT Journal Article
SR Electronic
T1 Substrate specificity profiling of SARS-CoV-2 main protease enables design of activity-based probes for patient-sample imaging
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.03.07.981928
DO 10.1101/2020.03.07.981928
A1 Wioletta Rut
A1 Katarzyna Groborz
A1 Linlin Zhang
A1 Xinyuanyuan Sun
A1 Mikolaj Zmudzinski
A1 Bartlomiej Pawlik
A1 Wojciech Młynarski
A1 Rolf Hilgenfeld
A1 Marcin Drag
YR 2020
UL http://biorxiv.org/content/early/2020/06/08/2020.03.07.981928.abstract
AB In December 2019, the first cases of infection with a novel coronavirus, SARS-CoV-2, were diagnosed in Wuhan, China. Due to international travel and human-to-human transmission, the virus spread rapidly inside and outside of China. Currently, there is no effective antiviral treatment for coronavirus disease 2019 (COVID-19); therefore, research efforts are focused on the rapid development of vaccines and antiviral drugs. The SARS-CoV-2 main protease constitutes one of the most attractive antiviral drug targets. To address this emerging problem, we have synthesized a combinatorial library of fluorogenic substrates with glutamine in the P1 position. We used it to determine the substrate preferences of the SARS-CoV and SARS-CoV-2 main proteases, using natural and a large panel of unnatural amino acids. On the basis of these findings, we designed and synthesized an inhibitor and two activity-based probes, for one of which we determined the crystal structure of its complex with the SARS-CoV-2 Mpro. Using this approach we visualized SARS-CoV-2 active Mpro within nasopharyngeal epithelial cells of a patient with active COVID-19 infection. The results of our work provide a structural framework for the design of inhibitors as antiviral agents or diagnostic tests.Competing Interest StatementWroclaw University of Science and Technology has filed a patent application covering compounds: Ac-Abu-Tle-Leu-Gln-VS, Biotin-PEG(4)-Abu-Tle-Leu-Gln-VS and Cy5-PEG(4)-Abu-Tle-Leu-Gln-VS as well as related compounds with W.R. and M.D. as inventors.Abu2-aminobutanoic acid;2-Abz2-(amino)benzoic acid;3-Abz3-(amino)benzoic acid;ACC7-amino-4-carbamoylmethylcoumarin;Dab2,4-diaminobutyric acid;Dhtdihydrotryptophan;HyCoSuLHybrid Combinatorial Substrate Library;Ornornithine;RFUrelative fluorescence unit;D-PhgD-phenylglycine;Thzthiazolidine-4-carboxylic acid;Tletert-leucine;