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http://hdl.handle.net/1942/40549
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DC Field | Value | Language |
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dc.contributor.author | Nikolaev, Andrey | - |
dc.contributor.author | Yudenko, Anna | - |
dc.contributor.author | SMOLENTSEVA, Anastasiia | - |
dc.contributor.author | Bogorodskiy, Andrey | - |
dc.contributor.author | Tsybrov, Fedor | - |
dc.contributor.author | Borshchevskiy, Valentin | - |
dc.contributor.author | Bukhalovich, Siarhei | - |
dc.contributor.author | V. Nazarenko, Vera | - |
dc.contributor.author | Kuznetsova, Elizaveta | - |
dc.contributor.author | Semenov, Oleg | - |
dc.contributor.author | Remeeva, Alina | - |
dc.contributor.author | Gushchin, Ivan | - |
dc.date.accessioned | 2023-07-06T07:23:31Z | - |
dc.date.available | 2023-07-06T07:23:31Z | - |
dc.date.issued | 2023 | - |
dc.date.submitted | 2023-07-03T13:49:49Z | - |
dc.identifier.citation | JOURNAL OF BIOLOGICAL CHEMISTRY, 299 (3) (Art N° 102977) | - |
dc.identifier.issn | - | |
dc.identifier.uri | http://hdl.handle.net/1942/40549 | - |
dc.description.abstract | Flavin-binding fluorescent proteins are promising geneti-cally encoded tags for microscopy. However, spectral properties of their chromophores (riboflavin, flavin mononucleotide, and flavin adenine dinucleotide) are notoriously similar even be-tween different protein families, which limits applications of flavoproteins in multicolor imaging. Here, we present a palette of 22 finely tuned fluorescent tags based on the thermostable LOV domain from Chloroflexus aggregans. We performed site saturation mutagenesis of three amino acid positions in the flavin-binding pocket, including the photoactive cysteine, to obtain variants with fluorescence emission maxima uniformly covering the wavelength range from 486 to 512 nm. We demonstrate three-color imaging based on spectral separation and two-color fluorescence lifetime imaging of bacteria, as well as two-color imaging of mammalian cells (HEK293T), using the proteins from the palette. These results highlight the possibility of fine spectral tuning of flavoproteins and pave the way for further applications of flavin-binding fluorescent proteins in fluorescence microscopy. | - |
dc.description.sponsorship | Generation and optical characterization of CagFbFP variants was supported by the Russian Science Foundation, grant number 21-64-00018. A. B. acknowledges the support of the Ministry of Science and Higher Education of the Russian Federation (agreement 075-03-2023-106, project FSMG-2020-0003) for fluorescence microscopy experiments. | - |
dc.language.iso | en | - |
dc.publisher | ELSEVIER | - |
dc.rights | 2023 THE AUTHORS. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/ | - |
dc.subject.other | UV-Vis spectroscopy | - |
dc.subject.other | flavin | - |
dc.subject.other | fluorescence | - |
dc.subject.other | microscopy | - |
dc.subject.other | protein engineering | - |
dc.subject.other | Humans | - |
dc.subject.other | Flavin Mononucleotide | - |
dc.subject.other | Flavin-Adenine Dinucleotide | - |
dc.subject.other | HEK293 Cells | - |
dc.subject.other | Flavoproteins | - |
dc.subject.other | Riboflavin | - |
dc.subject.other | Luminescent Proteins | - |
dc.title | Fine spectral tuning of a flavin-binding fluorescent protein for multicolor imaging | - |
dc.type | Journal Contribution | - |
dc.identifier.issue | 3 | - |
dc.identifier.volume | 299 | - |
local.format.pages | 11 | - |
local.bibliographicCitation.jcat | A1 | - |
dc.description.notes | Gushchin, I (corresponding author), Moscow Inst Phys & Technol, Res Ctr Mol Mech Aging & Age Related Dis, Dolgoprudnyi, Russia. | - |
dc.description.notes | ivan.gushchin@phystech.edu | - |
local.publisher.place | RADARWEG 29, 1043 NX AMSTERDAM, NETHERLANDS | - |
local.type.refereed | Refereed | - |
local.type.specified | Article | - |
local.bibliographicCitation.artnr | 102977 | - |
dc.identifier.doi | 10.1016/j.jbc.2023.102977 | - |
dc.identifier.pmid | 36738792 | - |
dc.identifier.isi | 001007053600001 | - |
dc.contributor.orcid | Semenov, Oleg/0000-0001-9470-0013; Nikolaev, Andrey/0009-0004-2479-5124; | - |
dc.contributor.orcid | Borshchevskiy, Valentin/0000-0003-4398-9712 | - |
local.provider.type | wosris | - |
local.description.affiliation | [Nikolaev, Andrey; Yudenko, Anna; Smolentseva, Anastasia; Bogorodskiy, Andrey; Tsybrov, Fedor; Borshchevskiy, Valentin; Bukhalovich, Siarhei; V. Nazarenko, Vera; Kuznetsova, Elizaveta; Semenov, Oleg; Remeeva, Alina; Gushchin, Ivan] Moscow Inst Phys & Technol, Res Ctr Mol Mech Aging & Age Related Dis, Dolgoprudnyi, Russia. | - |
local.description.affiliation | [Smolentseva, Anastasia] Hasselt Univ, Adv Opt Microscopy Ctr, Dynam Bioimaging Lab, Diepenbeek, Belgium. | - |
local.description.affiliation | [Smolentseva, Anastasia] Hasselt Univ, Biomed Res Inst, Diepenbeek, Belgium. | - |
local.uhasselt.international | yes | - |
item.fullcitation | Nikolaev, Andrey; Yudenko, Anna; SMOLENTSEVA, Anastasiia; Bogorodskiy, Andrey; Tsybrov, Fedor; Borshchevskiy, Valentin; Bukhalovich, Siarhei; V. Nazarenko, Vera; Kuznetsova, Elizaveta; Semenov, Oleg; Remeeva, Alina & Gushchin, Ivan (2023) Fine spectral tuning of a flavin-binding fluorescent protein for multicolor imaging. In: JOURNAL OF BIOLOGICAL CHEMISTRY, 299 (3) (Art N° 102977). | - |
item.fulltext | With Fulltext | - |
item.contributor | Nikolaev, Andrey | - |
item.contributor | Yudenko, Anna | - |
item.contributor | SMOLENTSEVA, Anastasiia | - |
item.contributor | Bogorodskiy, Andrey | - |
item.contributor | Tsybrov, Fedor | - |
item.contributor | Borshchevskiy, Valentin | - |
item.contributor | Bukhalovich, Siarhei | - |
item.contributor | V. Nazarenko, Vera | - |
item.contributor | Kuznetsova, Elizaveta | - |
item.contributor | Semenov, Oleg | - |
item.contributor | Remeeva, Alina | - |
item.contributor | Gushchin, Ivan | - |
item.accessRights | Open Access | - |
crisitem.journal.eissn | 1083-351X | - |
Appears in Collections: | Research publications |
Files in This Item:
File | Description | Size | Format | |
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Fine spectral tuning of a flavin-binding fluorescent protein for multicolor imaging.pdf | Published version | 2.63 MB | Adobe PDF | View/Open |
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