Identification of a Heme Activation Site on the MD-2/TLR4 Complex.
LPS
MD-2
NF-κB
TLR4
heme
Journal
Frontiers in immunology
ISSN: 1664-3224
Titre abrégé: Front Immunol
Pays: Switzerland
ID NLM: 101560960
Informations de publication
Date de publication:
2020
2020
Historique:
received:
06
02
2020
accepted:
28
05
2020
entrez:
23
7
2020
pubmed:
23
7
2020
medline:
7
4
2021
Statut:
epublish
Résumé
Myeloid differentiation factor-2 (MD-2) binds lipopolysaccharide (LPS) and initiates toll-like receptor-4 (TLR4) pro-inflammatory signaling. Heme also activates TLR4 signaling, but it is unknown if heme interacts with MD-2. Therefore, we examined MD-2 for a potential heme activation site. Heme-agarose and biotin-heme/streptavidin-agarose pulled down recombinant MD-2, which was inhibited by excess free heme. UV/visible spectroscopy confirmed MD-2-heme binding. To determine whether MD-2 was required for heme-mediated TLR4 signaling, HEK293 cells were transfected with MD-2, TLR4, CD14, and an NF-κB luciferase reporter, and then stimulated with heme or LPS. Heme or LPS treatment elicited robust reporter activity. Absence of MD-2, TLR4 or CD14 plasmid abolished NF-κB reporter responses to heme or LPS.
Identifiants
pubmed: 32695117
doi: 10.3389/fimmu.2020.01370
pmc: PMC7338675
doi:
Substances chimiques
LY96 protein, human
0
Lymphocyte Antigen 96
0
TLR4 protein, human
0
Toll-Like Receptor 4
0
Heme
42VZT0U6YR
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
1370Subventions
Organisme : NCI NIH HHS
ID : P30 CA077598
Pays : United States
Organisme : NIDDK NIH HHS
ID : R01 DK119167
Pays : United States
Organisme : NHLBI NIH HHS
ID : R01 HL114567
Pays : United States
Organisme : NHLBI NIH HHS
ID : T32 HL007062
Pays : United States
Informations de copyright
Copyright © 2020 Belcher, Zhang, Nguyen, Kiser, Nath, Hu, Trent and Vercellotti.
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