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Self-assembling as regular nanoparticles dramatically minimizes photobleaching of tumour-targeted GFP
Unzueta Elorza, Ugutz (Institut d'Investigació Biomèdica Sant Pau)
Roldán, Mònica (Institut de Recerca Sant Joan de Déu)
Pesarrodona Roches, Mireia (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Benítez, Raúl (Institut de Recerca Sant Joan de Déu)
Sánchez Chardi, Alejandro (Universitat de Barcelona. Departament de Biologia Evolutiva, Ecologia i Ciències Ambientals)
Conchillo-Solé, Oscar (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Mangues, Ramon 1957- (Institut d'Investigació Biomèdica Sant Pau)
Villaverde Corrales, Antonio (Universitat Autònoma de Barcelona. Departament de Genètica i de Microbiologia)
Vázquez Gómez, Esther (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")

Date: 2020
Abstract: Fluorescent proteins are useful imaging and theranostic agents, but their potential superiority over alternative dyes is weakened by substantial photobleaching under irradiation. Enhancing protein photostability has been attempted through diverse strategies, with irregular results and limited applicability. In this context, we wondered if the controlled oligomerization of Green Fluorescent Protein (GFP) as nanoscale supramolecular complexes could stabilize the fluorophore through the newly formed protein-protein contacts, and thus, enhance its global photostability. For that, we have here analyzed the photobleaching profile of several GFP versions, engineered to self-assemble as tumour-homing nanoparticles with different targeting, size and structural stability. This has been done under prolonged irradiation in confocal laser scanning microscopy and by small-angle X-ray scattering. The results show that the oligomerization of GFP at the nanoscale enhances, by more than seven-fold, the stability of fluorescence emission. Interestingly, GFP nanoparticles are much more resistant to X-ray damage than the building block counterparts, indicating that the gained photostability is linked to enhanced structural resistance to radiation. Therefore, the controlled oligomerization of self-assembling fluorescent proteins as protein nanoparticles is a simple, versatile and powerful method to enhance their photostability for uses in precision imaging and therapy. Statement of significance: Fluorescent protein assembly into regular and highly symmetric nanoscale structures has been identified to confer enhanced structural stability against radiation stresses dramatically reducing their photobleaching. Being this the main bottleneck in the use of fluorescent proteins for imaging and theranostics, this protein architecture engineering principle appears as a powerful method to enhance their photostability for a broad applicability in precision imaging, drug delivery and theranostics.
Grants: Ministerio de Ciencia e Innovación SAF2017-8819-C3
Ministerio de Ciencia e Innovación BIO2016-76063-R
Instituto de Salud Carlos III PI15/00272
Agència de Gestió d'Ajuts Universitaris i de Recerca 2017/SGR-229
Rights: Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, i la comunicació pública de l'obra, sempre que no sigui amb finalitats comercials, i sempre que es reconegui l'autoria de l'obra original. No es permet la creació d'obres derivades. Creative Commons
Language: Anglès
Document: Article ; recerca ; Versió acceptada per publicar
Subject: Nanoparticles ; Fluorescent proteins ; Photostability ; Self-assembling ; Tumour targeting
Published in: Acta Biomaterialia, Vol. 103 (Feb. 2020) , p. 272-280, ISSN 1878-7568

DOI: 10.1016/j.actbio.2019.12.003
PMID: 31812843


Postprint
35 p, 893.9 KB

The record appears in these collections:
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut de Biotecnologia i de Biomedicina (IBB)
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut de Recerca Sant Pau
Articles > Research articles
Articles > Published articles

 Record created 2020-11-09, last modified 2023-11-29



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