The Effects of the Combined Co-Expression of GroEL/ES and Trigger Factor Chaperones on Orthopoxvirus Phospholipase F13 Production in E. coli .

Publisher: MDPI AG Country of Publication: Switzerland NLM ID: 9918383086206676 Publication Model: Electronic Cited Medium: Internet ISSN: 2673-6284 (Electronic) Linking ISSN: 26736284 NLM ISO Abbreviation: BioTech (Basel) Subsets: PubMed not MEDLINE

Original Publication: Basel, Switzerland : MDPI AG, [2020]-

Heterologous protein expression often faces significant challenges, particularly when the target protein has posttranslational modifications, is toxic, or is prone to misfolding. These issues can result in low expression levels, aggregation, or even cell death. Such problems are exemplified by the expression of phospholipase p37, a critical target for chemotherapeutic drugs against pathogenic human orthopoxviruses, including monkeypox and smallpox viruses. The complex structure and broad enzymatic activity of phospholipase p37 render it toxic to host cells, necessitating specialized strategies for heterologous expression. In our study, we addressed these challenges using the vaccinia virus F13 protein as a model. We demonstrated that p37 can be effectively synthesized in E. coli as a GST fusion protein by co-expressing it with the GroEL/ES chaperone system and Trigger Factor chaperone.

Protein Sci. 1996 May;5(5):914-22. (PMID: 8732763)
N Engl J Med. 2022 Aug 25;387(8):679-691. (PMID: 35866746)
Microb Cell Fact. 2021 Oct 30;20(1):208. (PMID: 34717620)
ChemMedChem. 2022 Jun 20;17(12):e202100771. (PMID: 35388614)
Nat Methods. 2022 Jun;19(6):679-682. (PMID: 35637307)
Appl Microbiol Biotechnol. 2012 Feb;93(3):1065-74. (PMID: 21975693)
J Virol. 2016 Sep 12;90(19):8891-905. (PMID: 27466413)
Biotechnol Adv. 2024 Oct;75:108417. (PMID: 39038691)
Protein Expr Purif. 2022 Aug;195-196:106097. (PMID: 35470011)
Protein J. 2021 Oct;40(5):756-764. (PMID: 34052952)
J Virol. 1999 Feb;73(2):1108-17. (PMID: 9882312)
Viruses. 2023 Feb 19;15(2):. (PMID: 36851785)
J Biol Chem. 1997 Dec 19;272(51):32042-9. (PMID: 9405398)
Int J Antimicrob Agents. 2024 Feb;63(2):107068. (PMID: 38141836)
J Virol. 2018 May 14;92(11):. (PMID: 29540596)
Eur J Med Chem. 2021 Oct 5;221:113485. (PMID: 33965861)
Viruses. 2022 Dec 21;15(1):. (PMID: 36680072)
Virol J. 2009 Apr 28;6:44. (PMID: 19400954)
Future Virol. 2011 May;6(5):653-671. (PMID: 21837250)
Nature. 1987 Jul 30-Aug 5;328(6129):378-9. (PMID: 3112578)
Sci Rep. 2021 Sep 14;11(1):18241. (PMID: 34521893)
Nat Commun. 2022 Jul 15;13(1):4126. (PMID: 35840586)
Nature. 2021 Aug;596(7873):583-589. (PMID: 34265844)
Viruses. 2023 Jan 11;15(1):. (PMID: 36680245)
Antimicrob Agents Chemother. 2020 Mar 24;64(4):. (PMID: 31932370)
J Gen Virol. 2002 Dec;83(Pt 12):2915-2931. (PMID: 12466468)
Methods Mol Biol. 2011;681:259-80. (PMID: 20978970)
Microorganisms. 2023 Nov 06;11(11):. (PMID: 38004725)
J Clin Virol. 2024 Apr;171:105662. (PMID: 38432097)
Enzyme Microb Technol. 2000 Mar 1;26(5-6):451-458. (PMID: 10713220)
Appl Environ Microbiol. 2000 Mar;66(3):884-9. (PMID: 10698746)

No. 075-15-2021-1355 dated October 12, 2021 The study was supported by the Ministry of Science and Higher Education of the Russian Federation as a part of the implementation of certain activities of the Federal Scientific and Technical Program of the Development of Synchrotron and Neutron Research

Keywords: GroEL/ES; NIOCH-14; ST-246; chaperone; monkeypox virus; orthopoxviruses; p37; phospholipase F13; trigger factor

Date Created: 20241227 Latest Revision: 20250104

20250114

PMC11674265

10.3390/biotech13040057

39727494