LOV2-Controlled Photoactivation of Protein Trans-Splicing

Methods Mol Biol. 2017:1495:227-237. doi: 10.1007/978-1-4939-6451-2_15.

Abstract

Protein trans-splicing is a posttranslational modification that joins two protein fragments together via a peptide a bond in a process that does not require exogenous cofactors. Towards achieving cellular control, synthetically engineered systems have used a variety of stimuli such as small molecules and light. Recently, split inteins have been engineered to be photoactive by the LOV2 domain (named LOVInC). Herein, we discuss (1) designing of LOV2-activated target proteins (e.g., inteins), (2) selecting feasible splice sites for the extein, and (3) imaging cells that express LOVInC-based target exteins.

Keywords: Fluorescence; Inteins; Light-oxygen-voltage domain; Photoactivation; Protein trans-splicing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis Proteins* / biosynthesis
  • Arabidopsis Proteins* / chemistry
  • Arabidopsis Proteins* / genetics
  • DNA-Binding Proteins* / biosynthesis
  • DNA-Binding Proteins* / chemistry
  • DNA-Binding Proteins* / genetics
  • HeLa Cells
  • Humans
  • Inteins*
  • Photochemical Processes*
  • Protein Domains
  • Protein Splicing*
  • Recombinant Fusion Proteins* / biosynthesis
  • Recombinant Fusion Proteins* / chemistry
  • Recombinant Fusion Proteins* / genetics

Substances

  • Arabidopsis Proteins
  • DNA-Binding Proteins
  • LOV2 protein, Arabidopsis
  • Recombinant Fusion Proteins