A live cell hormone-binding assay on transgenic bacteria expressing a eukaryotic receptor protein

Anal Biochem. 2005 Dec 1;347(1):129-34. doi: 10.1016/j.ab.2005.09.012. Epub 2005 Sep 30.

Abstract

The investigation of hormone-receptor interaction normally needs isolation and extensive purification of the receptor protein or a particular receptor-containing fraction. To bypass these time- and resource-consuming procedures, we have established a live cell-based assay using transgenic bacteria expressing single eukaryotic receptors. Here we describe some biochemical features of the Arabidopsis cytokinin receptor CRE1/AHK4 expressed in Escherichia coli. The data show that the main characteristics of the ligand-receptor interaction, including binding affinity and ligand specificity, can be determined using intact bacteria expressing a functional receptor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis Proteins / analysis*
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Binding, Competitive
  • Cytokinins / metabolism*
  • Escherichia coli / genetics*
  • Ligands
  • Protein Kinases / analysis*
  • Protein Kinases / genetics
  • Protein Kinases / metabolism*
  • Receptors, Cell Surface / analysis*
  • Receptors, Cell Surface / genetics
  • Receptors, Cell Surface / metabolism*

Substances

  • Arabidopsis Proteins
  • Cytokinins
  • Ligands
  • Receptors, Cell Surface
  • Protein Kinases
  • WOL protein, Arabidopsis