Статья

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

I. Dyakov, D. Mavletova, I. Chernyshova, N. Snegireva, M. Gavrilova, K. Bushkova, M. Dyachkova, M. Alekseeva, V. Danilenko,
2021

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. © 2020 Elsevier Ltd

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  • 1. Version of Record от 2021-04-27

Метаданные

Об авторах
  • I. Dyakov
    I.I, Mechnikov Research Institute for Vaccines and Sera, Malyj Kazennyj Per., 5, Moscow, 105064, Russian Federation
  • D. Mavletova
    Vavilov Institute of General Genetics, Russian Academy of Sciences, Gubkina St., 3, Moscow, 119991, Russian Federation
  • I. Chernyshova
    Peoples’ Friendship University of Russia (RUDN University), Faculty of Ecology, International Institute for Strategic Development of Sectoral Economics, Miklukho-Maklaya St., 6, Moscow, 117198, Russian Federation
  • N. Snegireva
    Pharmabiotics Limited Liability Company, Bolshoy Boulevard, 42, Bldg. 1, 1238, Moscow, 121205, Russian Federation
  • M. Gavrilova
  • K. Bushkova
  • M. Dyachkova
  • M. Alekseeva
  • V. Danilenko
Название журнала
  • Anaerobe
Том
  • 65
Страницы
  • -
Ключевые слова
  • bacterial protein; cytokine; fibronectin; FN3 protein; interleukin 10; interleukin 1beta; interleukin 6; tumor necrosis factor; unclassified drug; Article; bacterial gene; bacterial strain; Bifidobacterium longum subsp. longum; controlled study; Escherichia coli; expression vector; fn3 gene; immunological procedures; in vitro study; molecular cloning; nonhuman; priority journal; protein binding; protein domain; protein expression; protein interaction; protein purification; sandwich ELISA
Издатель
  • Academic Press
Тип документа
  • journal article
Тип лицензии Creative Commons
  • CC
Правовой статус документа
  • Свободная лицензия
Источник
  • scopus