tdLanYFP, a yellow, bright, photostable and pH insensitive fluorescent protein for live cell imaging and FRET-based sensing strategies - Institut Génétique et Développement de Rennes Accéder directement au contenu
Pré-Publication, Document De Travail Année : 2021

tdLanYFP, a yellow, bright, photostable and pH insensitive fluorescent protein for live cell imaging and FRET-based sensing strategies

Yasmina Bousmah
  • Fonction : Auteur
Hana Valenta
Utkarsh Singh
  • Fonction : Auteur
Valérie Nicolas
  • Fonction : Auteur
Hélène Pasquier
Marc Tramier
Fabienne Merola
  • Fonction : Auteur
Marie Erard

Résumé

Abstract Yellow fluorescent proteins (YFP) are widely used as optical reporters in Förster Resonance Energy Transfer (FRET) based biosensors. Although great improvements have been done, the sensitivity of the biosensors is still limited by the low photostability and the poor fluorescence performances of YFPs at acidic pHs. Here, we characterize the yellow fluorescent protein, tdLanYFP, derived from the tetrameric protein from the cephalochordate B. lanceolatum , LanYFP. With a quantum yield of 0.92 and an extinction coefficient of 133 000 mol −1 .L.cm −1 , it is, to our knowledge, the brightest dimeric fluorescent protein available. Contrasting with EYFP and its derivatives, tdLanYFP has a very high photostability in vitro and in live cells. As a consequence, tdLanYFP allows imaging of cellular structures with sub-diffraction resolution using STED nanoscopy and is compatible with the use of spectro-microscopies in single molecule regimes. Its very low pK 1/2 of 3.9 makes tdLanYFP an excellent tag even at acidic pHs. Finally, we show that tdLanYFP is valuable FRET partner either as donor or acceptor in different biosensing modalities. Altogether, these assets make tdLanYFP a very attractive yellow fluorescent protein for long-term or single-molecule live cell imaging including FRET experiments at acidic pH.
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Dates et versions

hal-03383692 , version 1 (18-10-2021)

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Paternité - Pas d'utilisation commerciale - Pas de modification

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Yasmina Bousmah, Hana Valenta, Giulia Bertolin, Utkarsh Singh, Valérie Nicolas, et al.. tdLanYFP, a yellow, bright, photostable and pH insensitive fluorescent protein for live cell imaging and FRET-based sensing strategies. 2021. ⟨hal-03383692⟩
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