FN3 protein fragment containing two type III fibronectin domains from B. longum GT15 binds to human tumor necrosis factor alpha in vitro.
Bacterial Proteins
/ chemistry
Bifidobacteriales Infections
/ metabolism
Bifidobacterium
/ genetics
Computational Biology
/ methods
Cytokines
/ metabolism
Fibronectin Type III Domain
Fibronectins
/ chemistry
Host-Pathogen Interactions
Humans
Multigene Family
Protein Binding
Recombinant Proteins
/ genetics
Tumor Necrosis Factor-alpha
/ metabolism
Analysis
Bifidobacterium
Cytokines
Immunoferment
PFNA cluster
Receptors
fn3 gene
Journal
Anaerobe
ISSN: 1095-8274
Titre abrégé: Anaerobe
Pays: England
ID NLM: 9505216
Informations de publication
Date de publication:
Oct 2020
Oct 2020
Historique:
received:
24
02
2020
revised:
09
07
2020
accepted:
21
07
2020
pubmed:
11
8
2020
medline:
23
6
2021
entrez:
11
8
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 32771620
pii: S1075-9964(20)30103-7
doi: 10.1016/j.anaerobe.2020.102247
pmc: PMC7409735
pii:
doi:
Substances chimiques
Bacterial Proteins
0
Cytokines
0
Fibronectins
0
Recombinant Proteins
0
Tumor Necrosis Factor-alpha
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
102247Informations de copyright
Copyright © 2020 Elsevier Ltd. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of competing interest The authors declare no conflict of interest.
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