Characterization of an anti-FLAG antibody binding protein in V. cholerae.
Affinity Labels
/ metabolism
Amino Acid Sequence
Animals
Antibodies, Monoclonal
/ metabolism
Bacterial Proteins
/ genetics
Carrier Proteins
/ genetics
Escherichia coli
/ genetics
Mice
Mutation
Oligopeptides
/ immunology
Porins
/ genetics
Protein Binding
Recombinant Fusion Proteins
/ genetics
Vibrio cholerae
/ genetics
3xFLAG tag
Affinity tag
Epitope tag
Monoclonal anti-FLAG M2 antibody and Porin4
Non-specific binding
Journal
Biochemical and biophysical research communications
ISSN: 1090-2104
Titre abrégé: Biochem Biophys Res Commun
Pays: United States
ID NLM: 0372516
Informations de publication
Date de publication:
30 07 2020
30 07 2020
Historique:
received:
27
04
2020
accepted:
24
05
2020
pubmed:
9
6
2020
medline:
9
2
2021
entrez:
8
6
2020
Statut:
ppublish
Résumé
FLAG-tags are commonly used for protein abundance measurements and for identification of protein-protein interactions in living cells. We have observed that the cholera pathogen Vibrio cholerae encodes a FLAG-antibody-reactive protein and identified this protein as an outer membrane porin, Porin4, which contains a sequence very similar to the 3xFLAG epitope tag. We have demonstrated the binding affinity of the conserved peptide sequence (called Porin 4 tag) in Porin4 against monoclonal anti-FLAG M2 antibody. In addition, we created a porin4 deletion mutant, which can be used for background-less FLAG antibody detection experiments.
Identifiants
pubmed: 32505345
pii: S0006-291X(20)31106-2
doi: 10.1016/j.bbrc.2020.05.169
pmc: PMC7357423
mid: NIHMS1600899
pii:
doi:
Substances chimiques
Affinity Labels
0
Antibodies, Monoclonal
0
Bacterial Proteins
0
Carrier Proteins
0
Oligopeptides
0
Porins
0
Recombinant Fusion Proteins
0
FLAG peptide
98849-88-8
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
493-498Subventions
Organisme : NIAID NIH HHS
ID : R01 AI143704
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM123018
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM130971
Pays : United States
Organisme : NICHD NIH HHS
ID : R01 HD095296
Pays : United States
Informations de copyright
Copyright © 2020 Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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