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Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells
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Genetic Code Expansion and Optoproteomics.

Yuting Chen1, Linjie Lu2, Shixin Ye2,3,4

  • 1Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University (ECNU), Shanghai, China.

The Yale Journal of Biology and Medicine
|December 21, 2017
PubMed
Summary
This summary is machine-generated.

Genetic code expansion (GCE) enables engineering light-sensitive proteins by incorporating unnatural amino acids. This optoproteomics approach allows precise investigation of protein functions and interactions.

Keywords:
Genetic code expansionion channelskinases interactionsoptochemical geneticsoptoproteomicsunnatural amino acids

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Nature utilizes photoreceptor proteins for light sensing and response.
  • Genetic code expansion (GCE) offers tools to engineer novel photoreceptor proteins.
  • Unnatural amino acids (Uaas) with light-sensitive moieties are key to this transformation.

Purpose of the Study:

  • To review principles of genetic editing for GCE.
  • To discuss design concepts for engineering light-sensitive proteins.
  • To introduce the concept of optoproteomics.

Main Methods:

  • Site-specific incorporation of Uaas into proteins.
  • Utilizing Uaas for photo-decaging, cross-linking, or isomerization.
  • Engineering proteins like Cas9 for light-activated functions.

Main Results:

  • Various proteins (ion channels, GPCRs, kinases, Cas9) have been rendered light-responsive.
  • Enabled discovery of dynamic protein conformational changes.
  • Facilitated screening of ligand-protein and protein-protein interactions.

Conclusions:

  • GCE and Uaas create novel light-sensitive proteins.
  • Optoproteomics combines optical methods with proteomics for biological investigation.
  • Engineered light-sensitive proteins have significant implications for disease research and drug discovery.