Large-Scale Recombinant Production of the SARS-CoV-2 Proteome for High-Throughput and Structural Biology Applications

被引：32

作者：

Altincekic, Nadide ^{[1
,2
]}

Korn, Sophie Marianne ^{[2
,3
]}

Qureshi, Nusrat Shahin ^{[1
,2
]}

Dujardin, Marie ^{[4
]}

Ninot-Pedrosa, Marti ^{[4
]}

Abele, Rupert ^{[5
]}

Saad, Marie Jose Abi ^{[6
]}

Alfano, Caterina ^{[7
]}

Almeida, Fabio C. L. ^{[8
,9
]}

Alshamleh, Islam ^{[1
,2
]}

de Amorim, Gisele Cardoso ^{[8
,10
]}

Anderson, Thomas K. ^{[11
]}

Anobom, Cristiane D. ^{[8
,12
]}

Anorma, Chelsea ^{[13
]}

Bains, Jasleen Kaur ^{[1
,2
]}

Bax, Adriaan ^{[14
]}

Blackledge, Martin ^{[15
]}

Blechar, Julius ^{[1
,2
]}

Bockmann, Anja ^{[4
]}

Brigandat, Louis ^{[4
]}

Bula, Anna ^{[16
]}

Buetikofer, Matthias ^{[6
]}

Camacho-Zarco, Aldo R. ^{[15
]}

Carlomagno, Teresa ^{[17
,18
,19
]}

Caruso, Icaro Putinhon ^{[8
,9
,20
]}

Ceylan, Betul ^{[1
,2
]}

Chaikuad, Apirat ^{[21
,22
]}

Chu, Feixia ^{[23
]}

Cole, Laura ^{[4
]}

Crosby, Marquise G. ^{[24
]}

de Jesus, Vanessa ^{[1
,2
]}

Dhamotharan, Karthikeyan ^{[2
,3
]}

Felli, Isabella C. ^{[25
,26
]}

Ferner, Jan ^{[1
,2
]}

Fleischmann, Yanick ^{[6
]}

Fogeron, Marie-Laure ^{[4
]}

Fourkiotis, Nikolaos K. ^{[27
]}

Fuks, Christin ^{[1
]}

Fuertig, Boris ^{[1
,2
]}

Gallo, Angelo ^{[27
]}

Gande, Santosh L. ^{[1
,2
]}

Gerez, Juan Atilio ^{[6
]}

Ghosh, Dhiman ^{[6
]}

Gomes-Neto, Francisco ^{[8
,28
]}

Gorbatyuk, Oksana ^{[29
]}

Guseva, Serafima ^{[15
]}

Hacker, Carolin ^{[30
]}

Haefner, Sabine ^{[31
]}

Hao, Bing ^{[29
]}

Hargittay, Bruno ^{[1
,2
]}

机构：

[1] Goethe Univ Frankfurt, Inst Organ Chem & Chem Biol, Frankfurt, Germany

[2] Goethe Univ Frankfurt, Ctr Biomol Magnet Resonance BMRZ, Frankfurt, Germany

[3] Goethe Univ Frankfurt, Inst Mol Biosci, Frankfurt, Germany

[4] Lyon Univ, CNRS, Mol Microbiol & Struct Biochem, UMR 5086, Lyon, France

[5] Goethe Univ Frankfurt, Inst Biochem, Frankfurt, Germany

[6] Swiss Fed Inst Technol, Lab Phys Chem, Swiss Fed Inst Technol, Zurich, Switzerland

[7] Fdn RiMED, Struct Biol & Biophys Unit, Palermo, Italy

[8] Univ Fed Rio de Janeiro, Natl Ctr Nucl Magnet Resonance CNRMN, CENABIO, Rio De Janeiro, Brazil

[9] Univ Fed Rio de Janeiro, Inst Med Biochem, Rio De Janeiro, Brazil

[10] Univ Fed Rio de Janeiro, Multidisciplinary Ctr Res Biol NUMPEX, Campus Duque Caxias, Duque De Caxias, RJ, Brazil

[11] Univ Wisconsin, Inst Mol Virol, Madison, WI 53706 USA

[12] Univ Fed Rio de Janeiro, Inst Chem, Rio De Janeiro, Brazil

[13] Univ Calif Irvine, Dept Chem, Irvine, CA 92717 USA

[14] NIDDK, LCP, NIH, Bethesda, MD 20892 USA

[15] Univ Grenoble Alpes, IBS, CEA, CNRS, Grenoble, France

[16] Latvian Inst Organ Synth, Riga, Latvia

[17] Leibniz Univ Hannover, BMWZ, Hannover, Germany

[18] Leibniz Univ Hannover, Inst Organ Chem, Hannover, Germany

[19] Helmholtz Ctr Infect Res, Grp NMR Based Struct Chem, Braunschweig, Germany

[20] Sao Paulo State Univ UNESP, Multiuser Ctr Biomol Innovat CMIB, Dept Phys, Sao Jose Do Rio Preto, Brazil

[21] Goethe Univ Frankfurt, Inst Pharmaceut Chem, Frankfurt, Germany

[22] Buchmann Inst Mol Life Sci, Struct Genom Consortium, Frankfurt, Germany

[23] Univ New Hampshire, Dept Mol Cellular & Biomed Sci, Durham, NH 03824 USA

[24] Univ Calif Irvine, Dept Mol Biol & Biochem, Irvine, CA USA

[25] Univ Florence, Magnet Resonance Ctr CERM, Sesto Fiorentino, Italy

[26] Univ Florence, Dept Chem Ugo Schiff, Sesto Fiorentino, Italy

[27] Univ Patras, Dept Pharm, Patras, Greece

[28] Oswaldo Cruz Fdn FIOCRUZ, Lab Toxinol, Rio De Janeiro, Brazil

[29] UConn Hlth, Dept Mol Biol & Biophys, Farmington, CT USA

[30] Signals GmbH & Co KG, Frankfurt, Germany

[31] Fritz Lipmann Inst FLI, Leibniz Inst Aging, Jena, Germany

[32] Latvian Biomed Res & Study Ctr, Riga, Latvia

[33] CSIC, Rocasolano Inst Phys Chem IQFR, Madrid, Spain

[34] Goethe Univ Frankfurt, Inst Biophys Chem, Frankfurt, Germany

[35] Karlsruhe Inst Technol, IBG 4, Karlsruhe, Germany

[36] Univ Wisconsin, Dept Biochem, 420 Henry Mall, Madison, WI 53705 USA

[37] Univ Wisconsin, Natl Magnet Resonance Facil Madison, Madison, WI USA

[38] Tech Univ Darmstadt, Dept Biol, Darmstadt, Germany

[39] Martin Luther Univ Halle Wittenberg, Charles Tanford Prot Ctr, Inst Biochem & Biotechnol, Halle, Germany

来源：

FRONTIERS IN MOLECULAR BIOSCIENCES | 2021年 / 8卷

基金：

欧洲研究理事会; 欧盟地平线“2020”; 瑞士国家科学基金会;

关键词：

COVID-19; SARS-CoV-2; nonstructural proteins; structural proteins; accessory proteins; intrinsically disordered region; cell-free protein synthesis; NMR spectroscopy; RESPIRATORY SYNDROME-CORONAVIRUS; MAGNETIC-RESONANCE STRUCTURE; MURINE HEPATITIS-VIRUS; HOST GENE-EXPRESSION; CELL-FREE EXPRESSION; SARS-CORONAVIRUS; NUCLEOCAPSID PROTEIN; NONSTRUCTURAL PROTEIN-3; CRYSTAL-STRUCTURE; UNIQUE DOMAIN;

D O I：

10.3389/fmolb.2021.653148

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The highly infectious disease COVID-19 caused by the Betacoronavirus SARS-CoV-2 poses a severe threat to humanity and demands the redirection of scientific efforts and criteria to organized research projects. The international COVID19-NMR consortium seeks to provide such new approaches by gathering scientific expertise worldwide. In particular, making available viral proteins and RNAs will pave the way to understanding the SARS-CoV-2 molecular components in detail. The research in COVID19-NMR and the resources provided through the consortium are fully disclosed to accelerate access and exploitation. NMR investigations of the viral molecular components are designated to provide the essential basis for further work, including macromolecular interaction studies and high-throughput drug screening. Here, we present the extensive catalog of a holistic SARS-CoV-2 protein preparation approach based on the consortium's collective efforts. We provide protocols for the large-scale production of more than 80% of all SARS-CoV-2 proteins or essential parts of them. Several of the proteins were produced in more than one laboratory, demonstrating the high interoperability between NMR groups worldwide. For the majority of proteins, we can produce isotope-labeled samples of HSQC-grade. Together with several NMR chemical shift assignments made publicly available on covid19-nmr.com, we here provide highly valuable resources for the production of SARS-CoV-2 proteins in isotope-labeled form.

引用

页数：25

共 118 条

[1] Novel β-barrel fold in the nuclear magnetic resonance structure of the replicase nonstructural protein 1 from the severe acute respiratory syndrome coronavirus [J].