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

[1] SARS-CoV-2 Main Protease Active Site Ligands in the Human Metabolome
Sardanelli, Anna Maria
Isgro, Camilla
Palese, Luigi Leonardo
MOLECULES, 2021, 26 (05):
[2] Drug development targeting SARS-CoV-2 main protease
Bulut, Haydar
GLOBAL HEALTH & MEDICINE, 2022, 4 (06): : 296 - 300
[3] Structural biology of SARS-CoV-1/SARS-CoV-2 main protease
Gao, Yunyun
Kaub, Johannes
Santoni, Gianluca
Pearce, Nicholas
Thorn, Andrea
CRYSTALLOGRAPHY REVIEWS, 2023, 29 (02) : 76 - 101
[4] Discovery of novel inhibitors of SARS-CoV-2 main protease
Zheng, Lei
Chen, Yanmei
Bao, Jingxiao
He, Liping
Dong, Suzhen
Qi, Yifei
Zhang, John Z. H.
JOURNAL OF BIOMOLECULAR STRUCTURE & DYNAMICS, 2022, 40 (23): : 12526 - 12534
[5] Fullerene derivatives as inhibitors of the SARS-CoV-2 main protease
Katagishi, Daiki
Yasuda, Daisuke
Takahashi, Kyoko
Nakamura, Shigeo
Mashino, Tadahiko
Ohe, Tomoyuki
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 2023, 80
[6] A new inactive conformation of SARS-CoV-2 main protease
Fornasier, Emanuele
Macchia, Maria Ludovica
Giachin, Gabriele
Sosic, Alice
Pavan, Matteo
Sturlese, Mattia
Salata, Cristiano
Moro, Stefano
Gatto, Barbara
Bellanda, Massimo
Battistutta, Roberto
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2022, 78 : 363 - 378
[7] A cyclic peptide inhibitor of the SARS-CoV-2 main protease
Kreutzer, Adam G.
Krumberger, Maj
Diessner, Elizabeth M.
Parrocha, Chelsea Marie T.
Morris, Michael A.
Guaglianone, Gretchen
Butts, Carter T.
Nowick, James S.
EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY, 2021, 221
[8] SARS-CoV-2 Main Protease Inhibitors That Leverage Unique Interactions with the Solvent Exposed S3 Site of the Enzyme
Blankenship, Lauren R.
Yang, Kai S.
Vulupala, Veerabhadra R.
Alugubelli, Yugendar R.
Khatua, Kaustav
Coleman, Demonta
Ma, Xinyu R.
Sankaran, Banumathi
Cho, Chia-Chuan D.
Ma, Yuying
Neuman, Benjamin W.
Xu, Shiqing
Liu, Wenshe Ray
ACS MEDICINAL CHEMISTRY LETTERS, 2024, 15 (06): : 950 - 957
[9] Computational analysis of the interactions between Ebselen and derivatives with the active site of the main protease from SARS-CoV-2
Rieder, Guilherme Schmitt
Nogara, Pablo Andrei
Omage, Folorunsho Bright
Duarte, Tamie
Dalla Corte, Cristiane Lenz
da Rocha, Joao Batista Teixeira
COMPUTATIONAL BIOLOGY AND CHEMISTRY, 2023, 107
[10] The SARS-CoV-2 main protease as drug target
Ullrich, Sven
Nitsche, Christoph
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 2020, 30 (17)

← 1 2 3 4 5 →