A live cell hormone-binding assay on transgenic bacteria expressing a eukaryotic receptor protein.
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.
Romanov
Georgy A
GA
Free University of Berlin, Institute of Biology/Applied Genetics, Germany. gar@ippras.ru
Spíchal
Lukás
L
Lomin
Sergey N
SN
Strnad
Miroslav
M
Schmülling
Thomas
T
D029681:Arabidopsis Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D001667:Binding, Competitive; D003583:Cytokinins / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D008024:Ligands; D011494:Protein Kinases / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*
D029681:Arabidopsis Proteins; D003583:Cytokinins; D008024:Ligands; D011956:Receptors, Cell Surface; D011494:Protein Kinases; C421783:WOL protein, Arabidopsis