FN3 protein fragment containing two type III fibronectin domains from B. longum GT15 binds to human tumor necrosis factor alpha in vitro.

Autor: Dyakov IN; I.I, Mechnikov Research Institute for Vaccines and Sera, Malyj Kazennyj Per., 5, Moscow, Russia, 105064., Mavletova DA; Vavilov Institute of General Genetics, Russian Academy of Sciences, Gubkina St., 3, Moscow, Russia, 119991., Chernyshova IN; I.I, Mechnikov Research Institute for Vaccines and Sera, Malyj Kazennyj Per., 5, Moscow, Russia, 105064., Snegireva NA; I.I, Mechnikov Research Institute for Vaccines and Sera, Malyj Kazennyj Per., 5, Moscow, Russia, 105064., Gavrilova MV; I.I, Mechnikov Research Institute for Vaccines and Sera, Malyj Kazennyj Per., 5, Moscow, Russia, 105064., Bushkova KK; I.I, Mechnikov Research Institute for Vaccines and Sera, Malyj Kazennyj Per., 5, Moscow, Russia, 105064., Dyachkova MS; Vavilov Institute of General Genetics, Russian Academy of Sciences, Gubkina St., 3, Moscow, Russia, 119991., Alekseeva MG; Vavilov Institute of General Genetics, Russian Academy of Sciences, Gubkina St., 3, Moscow, Russia, 119991., Danilenko VN; Vavilov Institute of General Genetics, Russian Academy of Sciences, Gubkina St., 3, Moscow, Russia, 119991; Peoples' Friendship University of Russia (RUDN University), Faculty of Ecology, International Institute for Strategic Development of Sectoral Economics, Miklukho-Maklaya St., 6, Moscow, Russia, 117198; Pharmabiotics Limited Liability Company, Bolshoy Boulevard, 42, Bldg. 1, 1238, Moscow, Russia, 121205. Electronic address: valerid@vigg.ru.
Jazyk: angličtina
Zdroj: Anaerobe [Anaerobe] 2020 Oct; Vol. 65, pp. 102247. Date of Electronic Publication: 2020 Aug 06.
DOI: 10.1016/j.anaerobe.2020.102247
Abstrakt: Most species of the genus Bifidobacterium contain the gene cluster PFNA, which is presumably involved in the species-specific communication between bacteria and their hosts. The gene cluster PFNA consists of five genes including fn3, which codes for a protein containing two fibronectin type III domains. Each fibronectin domain contains sites similar to cytokine-binding sites of human receptors. Based on this finding we assumed that this protein would bind specifically to human cytokines in vitro. We cloned a fragment of the fn3 gene (1503 bp; 501 aa) containing two fibronectin domains, from the strain B. longum subsp. longum GT15. After cloning the fragment into the expression vector pET16b and expressing it in E. coli, the protein product was purified to a homogenous state for further analysis. Using the immunoferment method, we tested the purified fragment's ability to bind the following human cytokines: IL-1β, IL-6, IL-10, TNFα. We developed a sandwich ELISA system to detect any specific interactions between the purified protein and any of the studied cytokines. We found that the purified protein fragment only binds to TNFα.
Competing Interests: Declaration of competing interest The authors declare no conflict of interest.
(Copyright © 2020 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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