Quantifying phagosomal HOCl at single immune-cell resolution.
DNA-based reporter
Fluorescent probe
Hypochlorous acid
Immune cells
Myeloperoxidase
Phagosome
pH
Journal
Methods in cell biology
ISSN: 0091-679X
Titre abrégé: Methods Cell Biol
Pays: United States
ID NLM: 0373334
Informations de publication
Date de publication:
2021
2021
Historique:
entrez:
6
7
2021
pubmed:
7
7
2021
medline:
26
11
2021
Statut:
ppublish
Résumé
Neutralization of pathogens by phagocytic immune cells requires the biogenesis of a compartmentalized hotspot of reactive species called the phagosome. One of these reactive species is hypochlorous acid (HOCl), produced by the enzyme myeloperoxidase (MPO) after the phagosome fuses with the lysosome. Mapping HOCl during phagosome maturation can report on pathogen killing and offer insights into regulation of MPO activity, mechanisms of resistance and host-pathogen interactions. However, this has been difficult because of a lack of a suitable method to chemically map a transient organelle with pH fluctuations like the phagosome. Here, we detail a protocol for quantifying HOCl dynamics in phagosomes using a fluorescent DNA-based reporter. Compared to traditional methods of visualizing HOCl or measuring MPO activity, this method offers sub-cellular spatial resolution and the capacity to assay HOCl production with single cell resolution.
Identifiants
pubmed: 34225911
pii: S0091-679X(20)30189-8
doi: 10.1016/bs.mcb.2020.10.006
pii:
doi:
Substances chimiques
Hypochlorous Acid
712K4CDC10
Peroxidase
EC 1.11.1.7
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
119-136Subventions
Organisme : NINDS NIH HHS
ID : R01 NS112139
Pays : United States
Organisme : NINDS NIH HHS
ID : R21 NS114428
Pays : United States
Informations de copyright
Copyright © 2021 Elsevier Inc. All rights reserved.