Journal article
A rationally designed orthogonal synthetase for genetically encoded fluorescent amino acids
Heliyon, Vol.6(10), pp.e05140-e05140
10/2020
DOI: 10.1016/j.heliyon.2020.e05140
PMCID: PMC7550906
PMID: 33083608
Abstract
The incorporation of non-canonical amino acids into proteins has emerged as a promising strategy to manipulate and study protein structure-function relationships with superior precision in vitro and in vivo. To date, fluorescent non-canonical amino acids (f-ncAA) have been successfully incorporated in proteins expressed in bacterial systems, Xenopus oocytes, and HEK-293T cells. Here, we describe the rational generation of a novel orthogonal aminoacyl-tRNA synthetase based on the E. coli tyrosine synthetase that is capable of encoding the f-ncAA tyr-coumarin in HEK-293T cells.
Bioorganic Chemistry; Bioinformatics; Proteins; Biochemistry; Molecular Biology; Unnatural amino acids, aminoacyl-tRNA synthetase, coumarin, fluorescence.
Details
- Title: Subtitle
- A rationally designed orthogonal synthetase for genetically encoded fluorescent amino acids
- Creators
- Ximena Steinberg - Physiology Department, Faculty of Medicine, Universidad Austral de Chile, Campus Isla Teja, Valdivia, 5110566, ChileJason Galpin - Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA 52242, United StatesGibran Nasir - Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA 52242, United StatesRomina V Sepúlveda - Center for Bioinformatics and Integrative Biology, Facultad de Ciencias de la Vida, Universidad Andres Bello, Avenida Republica 330, Santiago, ChileErnesto Ladron de Guevara - Departamento de Fisiología, Facultad de Medicina, Universidad Nacional Autónoma de México, Distrito Federal 04510, MexicoFernando Gonzalez-Nilo - Center for Bioinformatics and Integrative Biology, Facultad de Ciencias de la Vida, Universidad Andres Bello, Avenida Republica 330, Santiago, ChileLeon D Islas - Departamento de Fisiología, Facultad de Medicina, Universidad Nacional Autónoma de México, Distrito Federal 04510, MexicoChristopher A Ahern - Department of Molecular Physiology and Biophysics, University of Iowa, Iowa City, IA 52242, United StatesSebastian E Brauchi - Physiology Department, Faculty of Medicine, Universidad Austral de Chile, Campus Isla Teja, Valdivia, 5110566, Chile
- Resource Type
- Journal article
- Publication Details
- Heliyon, Vol.6(10), pp.e05140-e05140
- Publisher
- Elsevier Ltd
- DOI
- 10.1016/j.heliyon.2020.e05140
- PMID
- 33083608
- PMCID
- PMC7550906
- ISSN
- 2405-8440
- eISSN
- 2405-8440
- Grant note
- name: DRI-CONICYT/CONACYT, award: PCCI12023; name: Iniciativa Cientifica Milenio (ANID; DOI: 10.13039/100000002, name: National Institutes of Health; name: FONDECYT, award: 1170733; name: DGAPA-PAPIIT-UNAM, award: IN209515 and IN203318; DOI: 10.13039/100000002, name: National Institutes of Health, award: GM106569, GM087519; name: FONDECYT, award: 1191868; name: Anillo Cientifico
- Language
- English
- Date published
- 10/2020
- Academic Unit
- Molecular Physiology and Biophysics; Iowa Neuroscience Institute
- Record Identifier
- 9984070753802771
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