3CLpro of SARS-CoV-2
with an N-terminal
MBP-tag, sensitive internally quenched fluorogenic substrate, and
assay buffer were obtained from BPS Bioscience (San Diego, CA, USA).
The enzyme was expressed in E. coli expression system,
and has a molecular weight of 77.5 kDa. The peptide substrate contains
a 14 amino sequence (KTSAVLQSGFRKME) with Dabcyl and Edans attached
on its N- and C-terminals, respectively. The reaction buffer is composed
of 20 mM Tris-HCl (pH 7.3), 100 mM NaCl, 1 mM EDTA, 0.01% BSA (bovine
serum albumin), and 1 mM 1,4-dithio-d ,l -threitol
(DTT). GC376 (CAS No: 1416992-39-6) was purchased from Aobious (Gloucester,
MA, USA). Library of Pharmacologically Active Compounds (LOPAC) was
purchased from Sigma-Aldrich (St. Louis, MO, USA). All other compound
libraries were sourced by the National Center for Advancing Translational
Sciences (NCATS) including the NCATS Pharmaceutical Collection (NPC),29 (link) anti-infective, MIPE5.0, and NPACT libraries.
The LOPAC library has 1280 compounds consisting of marketed drugs
and pharmaceutically relevant structures with biological activities.
The NPC library contains 2552 FDA approved drugs, investigational
drugs, animal drugs, and anti-infectives. The anti-infective library
is a NCATS collection that contains 739 compounds that specifically
target viruses. The MIPE 5.0 library includes 2480 compounds that
are mixed with approved and investigational compounds, and mechanistic
based compounds focusing on oncology. The NPACT library contains 5099
structurally diverse compounds consisting of approved drugs, investigational
drugs, and natural products.
with an N-terminal
MBP-tag, sensitive internally quenched fluorogenic substrate, and
assay buffer were obtained from BPS Bioscience (San Diego, CA, USA).
The enzyme was expressed in E. coli expression system,
and has a molecular weight of 77.5 kDa. The peptide substrate contains
a 14 amino sequence (KTSAVLQSGFRKME) with Dabcyl and Edans attached
on its N- and C-terminals, respectively. The reaction buffer is composed
of 20 mM Tris-HCl (pH 7.3), 100 mM NaCl, 1 mM EDTA, 0.01% BSA (bovine
serum albumin), and 1 mM 1,4-dithio-
(DTT). GC376 (CAS No: 1416992-39-6) was purchased from Aobious (Gloucester,
MA, USA). Library of Pharmacologically Active Compounds (LOPAC) was
purchased from Sigma-Aldrich (St. Louis, MO, USA). All other compound
libraries were sourced by the National Center for Advancing Translational
Sciences (NCATS) including the NCATS Pharmaceutical Collection (NPC),29 (link) anti-infective, MIPE5.0, and NPACT libraries.
The LOPAC library has 1280 compounds consisting of marketed drugs
and pharmaceutically relevant structures with biological activities.
The NPC library contains 2552 FDA approved drugs, investigational
drugs, animal drugs, and anti-infectives. The anti-infective library
is a NCATS collection that contains 739 compounds that specifically
target viruses. The MIPE 5.0 library includes 2480 compounds that
are mixed with approved and investigational compounds, and mechanistic
based compounds focusing on oncology. The NPACT library contains 5099
structurally diverse compounds consisting of approved drugs, investigational
drugs, and natural products.
