Structural and photophysical characterization of the small ultra-red fluorescent protein.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
12 07 2023
12 07 2023
Historique:
received:
27
06
2022
accepted:
29
06
2023
medline:
14
7
2023
pubmed:
13
7
2023
entrez:
12
7
2023
Statut:
epublish
Résumé
The small Ultra-Red Fluorescent Protein (smURFP) represents a new class of fluorescent protein with exceptional photostability and brightness derived from allophycocyanin in a previous directed evolution. Here, we report the smURFP crystal structure to better understand properties and enable further engineering of improved variants. We compare this structure to the structures of allophycocyanin and smURFP mutants to identify the structural origins of the molecular brightness. We then use a structure-guided approach to develop monomeric smURFP variants that fluoresce with phycocyanobilin but not biliverdin. Furthermore, we measure smURFP photophysical properties necessary for advanced imaging modalities, such as those relevant for two-photon, fluorescence lifetime, and single-molecule imaging. We observe that smURFP has the largest two-photon cross-section measured for a fluorescent protein, and that it produces more photons than organic dyes. Altogether, this study expands our understanding of the smURFP, which will inform future engineering toward optimal FPs compatible with whole organism studies.
Identifiants
pubmed: 37438348
doi: 10.1038/s41467-023-39776-9
pii: 10.1038/s41467-023-39776-9
pmc: PMC10338489
doi:
Substances chimiques
Luminescent Proteins
0
Biliverdine
O9MIA842K9
Coloring Agents
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Intramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
4155Subventions
Organisme : NCI NIH HHS
ID : 75N91019D00024
Pays : United States
Organisme : NIAID NIH HHS
ID : R01 AI096837
Pays : United States
Organisme : NINDS NIH HHS
ID : U01 NS090590
Pays : United States
Organisme : NIGMS NIH HHS
ID : R35 GM118067
Pays : United States
Organisme : NIGMS NIH HHS
ID : T32 GM007240
Pays : United States
Organisme : Intramural NIH HHS
ID : ZIA BC011627
Pays : United States
Organisme : NINDS NIH HHS
ID : U24 NS109107
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM086197
Pays : United States
Informations de copyright
© 2023. The Author(s).
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