The Fluorescence-Activating and Absorption-Shifting Tag (FAST) Enables Live-Cell Fluorescence Imaging of Methanococcus maripaludis.
FAST
archaea
fluorescence microscopy
Journal
Journal of bacteriology
ISSN: 1098-5530
Titre abrégé: J Bacteriol
Pays: United States
ID NLM: 2985120R
Informations de publication
Date de publication:
19 07 2022
19 07 2022
Historique:
pubmed:
4
6
2022
medline:
22
7
2022
entrez:
3
6
2022
Statut:
ppublish
Résumé
Live-cell fluorescence imaging of methanogenic archaea has been limited due to the strictly anoxic conditions required for growth and issues with autofluorescence associated with electron carriers in central metabolism. Here, we show that the fluorescence-activating and absorption-shifting tag (FAST) complexed with the fluorogenic ligand 4-hydroxy-3-methylbenzylidene-rhodanine (HMBR) overcomes these issues and displays robust fluorescence in Methanococcus maripaludis. We also describe a mechanism to visualize cells under anoxic conditions using a fluorescence microscope. Derivatives of FAST were successfully applied for protein abundance analysis, subcellular localization analysis, and determination of protein-protein interactions. FAST fusions to both formate dehydrogenase (Fdh) and F
Identifiants
pubmed: 35657707
doi: 10.1128/jb.00120-22
pmc: PMC9295556
doi:
Substances chimiques
Ligands
0
Formate Dehydrogenases
EC 1.17.1.9
Types de publication
Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0012022Subventions
Organisme : DOD | US Army | RDECOM | Army Research Office (ARO)
ID : W911NF-19-1-0024
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