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https://hdl.handle.net/10316/109788
Título: | An improved Ras sensor for highly sensitive and quantitative FRET-FLIM imaging | Autor: | Oliveira, Ana F. Yasuda, Ryohei |
Data: | 2013 | Editora: | Public Library of Science | Projeto: | National Institute of Health (R01MH080047, R01NS068410 Pre-Doctoral Fellowship from the Portuguese Foundation for Science and Technology (SFRH/BD/15226/2004) Young Investigator Award, Children’s Tumor Foundation (YIA # 2009-01-014) Howard Hughes Medical Institute |
Título da revista, periódico, livro ou evento: | PLoS ONE | Volume: | 8 | Número: | 1 | Resumo: | Ras is a signaling protein involved in a variety of cellular processes. Hence, studying Ras signaling with high spatiotemporal resolution is crucial to understanding the roles of Ras in many important cellular functions. Previously, fluorescence lifetime imaging (FLIM) of fluorescent resonance energy transfer (FRET)-based Ras activity sensors, FRas and FRas-F, have been demonstrated to be useful for measuring the spatiotemporal dynamics of Ras signaling in subcellular micro-compartments. However the predominantly nuclear localization of the sensors' acceptor has limited its sensitivity. Here, we have overcome this limitation and developed two variants of the existing FRas sensor with different affinities: FRas2-F (K(d)∼1.7 µM) and FRas2-M (K(d)∼0.5 µM). We demonstrate that, under 2-photon fluorescence lifetime imaging microscopy, FRas2 sensors provide higher sensitivity compared to previous sensors in 293T cells and neurons. | URI: | https://hdl.handle.net/10316/109788 | ISSN: | 1932-6203 | DOI: | 10.1371/journal.pone.0052874 | Direitos: | openAccess |
Aparece nas coleções: | I&D CNC - Artigos em Revistas Internacionais |
Ficheiros deste registo:
Ficheiro | Descrição | Tamanho | Formato | |
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An-Improved-Ras-Sensor-for-Highly-Sensitive-and-Quantitative-FRETFLIM-ImagingPLoS-ONE.pdf | 797.57 kB | Adobe PDF | Ver/Abrir |
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