Development of an active-site titrant for SARS-CoV-2 main protease as an indispensable tool for evaluating enzyme kinetics

被引：0

作者：

Voget, Rabea ^{[1
]}

Breidenbach, Julian ^{[1
]}

Claff, Tobias ^{[1
]}

Hingst, Alexandra ^{[1
]}

Sylvester, Katharina ^{[1
]}

Steinebach, Christian ^{[1
]}

Vu, Lan Phuong ^{[1
]}

Weisse, Renato H. ^{[2
]}

Bartz, Ulrike ^{[3
]}

Straeter, Norbert ^{[2
]}

Mueller, Christa E. ^{[1
]}

Guetschow, Michael ^{[1
]}

机构：

[1] Univ Bonn, Pharmaceut Inst, Pharmaceut & Med Chem, D-53121 Bonn, Germany

[2] Univ Leipzig, Inst Bioanalyt Chem, Ctr Biotechnol & Biomed, D-04103 Leipzig, Germany

[3] Univ Appl Sci Bonn Rhein Sieg, Dept Nat Sci, D-53359 Rheinbach, Germany

来源：

ACTA PHARMACEUTICA SINICA B | 2024年 / 14卷 / 05期

关键词：

COVID-19; SARS-CoV-2; Main protease; Peptide nitriles; Fluorogenic substrates; Active-site titration; X-ray crystallography; Inner filter effect; SUBSTRATE-INHIBITION; FLUORESCENCE;

D O I：

10.1016/j.apsb.2024.03.0012211-3835

中图分类号：

R9 [药学];

学科分类号：

1007 ;

摘要：

A titrant for the SARS-CoV-2 main protease (M-pro) was developed that enables, for the first time, the exact determination of the concentration of the enzymatically active M-pro by active-site titration. The covalent binding mode of the tetrapeptidic titrant was elucidated by the determination of the crystal structure of the enzyme-titrant complex. Four fluorogenic substrates of M-pro, including a prototypical, internally quenched Dabcyl-EDANS peptide, were compared in terms of solubility under typical assay conditions. By exploiting the new titrant, key kinetic parameters for the M-pro-catalyzed cleavage of these substrates were determined.

引用

页码：2349 / 2357

页数：9

共 50 条

[31] Insights into the Interaction Mechanism of Boceprevir with SARS-CoV-2 Main Protease
Kaur, Gurmeet
Goyal, Bhupesh
CHEMISTRYSELECT, 2023, 8 (28):
[32] A Genetic Trap in Yeast for Inhibitors of SARS-CoV-2 Main Protease
Alalam, Hanna
Sigurdardottir, Sunniva
Bourgard, Catarina
Tiukova, Ievgeniia
King, Ross D.
Grotli, Morten
Sunnerhagen, Per
MSYSTEMS, 2021, 6 (06)
[33] Structural insights into SARS-CoV-2 Main Protease maturation process
Noske, Gabriela Dias
Nakamura, Aline Minali
Gawriljuk, Victor Oliveira
Fernandes, Rafaela Sachetto
Godoy, Andre Schutzer
Oliva, Glaucius
BIOPHYSICAL REVIEWS, 2021, 13 (06) : 1372 - 1372
[34] A Crystallographic Snapshot of SARS-CoV-2 Main Protease Maturation Process
Noske, Gabriela D.
Nakamura, Aline Minali
Gawriljuk, Victor O.
Fernandes, Rafaela S.
Lima, Gustavo M. A.
Rosa, Higor V. D.
Pereira, Humberto D.
Zeri, Ana C. M.
Nascimento, Andrey F. Z.
Freire, Marjorie C. L. C.
Oliva, Glaucius
Godoy, Andre S.
ACTA CRYSTALLOGRAPHICA A-FOUNDATION AND ADVANCES, 2021, 77 : C34 - C34
[35] Cotton flower metabolites inhibit SARS-CoV-2 main protease
Zhang, Yufang
Li, Wenkang
Hu, Yiming
Ding, Tianze
Zafa, Mubashar
Jia, Xue
Zhang, Liya
Ren, Maozhi
Li, Fuguang
Wang, Wenjing
FEBS OPEN BIO, 2022, 12 (10): : 1886 - 1895
[36] Zinc thiotropolone combinations as inhibitors of the SARS-CoV-2 main protease
DeLaney, Christopher
Sheng, Yan
Pectol, D. Chase
Vantansever, Erol
Zhang, Hanyuan
Bhuvanesh, Nattamai
Salas, Isaiah
Liu, Wenshe R.
Fierke, Carol F.
Darensbourg, Marcetta Y.
DALTON TRANSACTIONS, 2021, 50 (35) : 12226 - 12233
[37] In silicoallicin inducedS-thioallylation of SARS-CoV-2 main protease
Shekh, Shamasoddin
Reddy, K. Kasi Amarnath
Gowd, Konkallu Hanumae
JOURNAL OF SULFUR CHEMISTRY, 2021, 42 (01) : 109 - 120
[38] Plant protease inhibitors against SARS-CoV-2 main protease: an in silico approach
Lima, Adrianne M.
de Souza, Adson A.
Amaral, Jackson L.
Freire, Valder N.
Souza, Pedro F.
de Oliveira, Hermogenes D.
FUTURE VIROLOGY, 2023, 18 (07) : 439 - 454
[39] Non-peptidic inhibitors targeting SARS-CoV-2 main protease: A review
Xiao, Ya-Qi
Long, Jiao
Zhang, Shuang-Shuang
Zhu, Yuan -Yuan
Gu, Shuang-Xi
BIOORGANIC CHEMISTRY, 2024, 147
[40] Targeting allosteric pockets of SARS-CoV-2 main protease Mpro
Bhat, Zahoor Ahmad
Chitara, Dheeraj
Iqbal, Jawed
Sanjeev, B. S.
Madhumalar, Arumugam
JOURNAL OF BIOMOLECULAR STRUCTURE & DYNAMICS, 2022, 40 (14) : 6603 - 6618

← 1 2 3 4 5 →