Experimental characterization of in silico red-shift-predicted iLOVL470T/Q489K and iLOVV392K/F410V/A426S mutants
Abstract: iLOV is a flavin mononucleotide-binding fluorescent protein used for in vivo cellular imaging similar to the green fluorescent protein. To expand the range of applications of iLOV, spectrally tuned red-shifted variants are desirable to reduce phototoxicity and allow for better tissue penetration. In this report, we experimentally tested two iLOV mutants, iLOVL470T/Q489K and iLOVV392K/F410V/A426S, which were previously computationally proposed by (Khrenova J. Phys. Chem. B 2017, 121 (43), pp 10018−10025) to have red-shifted excitation and emission spectra. While iLOVL470T/Q489K is about 20% brighter compared to the WT in vitro, it exhibits a blue shift in contrast to quantum mechanics/molecular mechanics (QM/MM) predictions. Additional optical characterization of an iLOVV392K mutant revealed that V392 is essential for cofactor binding and, accordingly, variants with V392K mutation are unable to bind to FMN. iLOVL470T/Q489K and iLOVV392K/F410V/A426S are expressed at low levels and have no detectable fluorescence in living cells, preventing their utilization in imaging applications
- Location
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Deutsche Nationalbibliothek Frankfurt am Main
- Extent
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Online-Ressource
- Language
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Englisch
- Notes
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ACS omega. - 7, 23 (2022) , 19555-19560, ISSN: 2470-1343
- Event
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Veröffentlichung
- (where)
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Freiburg
- (who)
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Universität
- (when)
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2022
- Creator
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Wehler, Pierre
Armbruster, Daniel
Günter, Andreas
Schleicher, Erik
Di Ventura, Barbara
Öztürk, Mehmet Ali
- Contributor
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Arbeitsgruppe: Biologische Signalforschung
- DOI
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10.1021/acsomega.2c01283
- URN
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urn:nbn:de:bsz:25-freidok-2278437
- Rights
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Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
- Last update
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15.08.2025, 7:36 AM CEST
Data provider
Deutsche Nationalbibliothek. If you have any questions about the object, please contact the data provider.
Associated
- Wehler, Pierre
- Armbruster, Daniel
- Günter, Andreas
- Schleicher, Erik
- Di Ventura, Barbara
- Öztürk, Mehmet Ali
- Arbeitsgruppe: Biologische Signalforschung
- Universität
Time of origin
- 2022
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