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Engineering ‘Golden’ Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
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1Institute of Chemistry L 1, Department of Biocatalysis,Technical University of Berlin, 2Institute of Chemistry PC 14, Department of Bioenergetics,Technical University of Berlin, 3Institute of Chemistry TC 7, Department of Physical Chemistry/Molecular Material Sciences,Technical University of Berlin
Chapitres
- 00:05Titre
- 01:46Recombinant Protein Expression
- 04:05Target Protein Purification via Immobilized Metal Ion Affinity Chromatography
- 05:20Intact Protein Mass Analysis by High-performance Liquid Chromatography Coupled to Electrospray Ionization Time-of-flight Mass Spectrometry
- 06:38Fluorescence Lifetime Measurements and Decay-associated Spectra of GdFP
- 07:59Results: Gold Fluorescent Protein Analysis by Mass Spectrometry and Fluorescence
- 10:13Conclusion
Synthetic biology enables the engineering of proteins with unprecedented properties using the co-translational insertion of non-canonical amino acids. Here, we presented how a spectrally red-shifted variant of a GFP-type fluorophore with novel fluorescence spectroscopic properties, termed "gold" fluorescent protein (GdFP), is produced in E. coli via selective pressure incorporation (SPI).
