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Optical Control of a Neuronal Protein Using a Genetically Encoded Unnatural Amino Acid in Neurons
Published on: March 28, 2016
Un aminoácido fotoenjaulado codificado genéticamente
Ning Wu1, Alexander Deiters, T Ashton Cropp
1Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Journal of the American Chemical Society
|November 4, 2004
Resumen
Los investigadores crearon un nuevo par de tRNA/sintetasa para la levadura, permitiendo la inserción de aminoácidos específicos en las proteínas utilizando el codón TAG. Esto hace avanzar la biología sintética y la ingeniería de proteínas.
Área de la Ciencia:
- La bioquímica es la bioquímica.
- Biología Molecular Biología Molecular
- Biología sintética Biología sintética.
Sus antecedentes:
- Los pares ortogonales de tRNA/sintetasa son herramientas cruciales para expandir el código genético.
- Los sistemas anteriores tienen limitaciones en la levadura, lo que requiere nuevos enfoques para la incorporación de aminoácidos no naturales.
Objetivo del estudio:
- Desarrollar un segundo par de ARN de transferencia ortogonal (ARNt) / sintetasa para su uso en levaduras.
- Para permitir la inserción específica del sitio de aminoácidos no naturales en las proteínas de la levadura.
Principales métodos:
- Utilizó el par Escherichia coli tRNALeu/leucyl tRNA-sintetasa como base para desarrollar el nuevo sistema.
- Empleó una nueva selección genética para identificar mutantes específicos de la sintetasa.
- Probado el sistema con ácido alfa-aminocaprílico y cisteína o-nitrobenzil.
Principales resultados:
- Desarrolló con éxito una segunda pareja ortogonal de tRNA/sintetasa funcional en levaduras.
- Se han identificado mutantes de la sintetasa capaces de cargar selectivamente un tRNA supresor del ámbar.
- Demostró la inserción de ácido alfa-aminocaprílico y cisteína o-nitrobenzil en proteínas en el codón TAG amber nonsense.
Conclusiones:
- El par tRNA/sintetasa desarrollado proporciona una nueva herramienta para incorporar aminoácidos no naturales a las proteínas de levadura.
- Este sistema expande las capacidades de expansión del código genético en los eucariotas.
- Facilita los avances en la ingeniería de proteínas y las aplicaciones de biología sintética.
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