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PMID: 16246292 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

Analytical biochemistry ·Vol. 347 ·No. 1 ·2005-12-01 ·Pages 129-34

Romanov GA, Spíchal L, Lomin SN, Strnad M, Schmülling T

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.

MeSH Terms
Arabidopsis Proteins/analysis,genetics,metabolism Binding, Competitive Cytokinins/metabolism Escherichia coli/genetics Ligands Protein Kinases/analysis,genetics,metabolism Receptors, Cell Surface/analysis,genetics,metabolism
Chemicals
Arabidopsis Proteins Cytokinins Ligands Receptors, Cell Surface Protein Kinases WOL protein, Arabidopsis
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Romanov Georgy A
Free University of Berlin, Institute of Biology/Applied Genetics, Germany. [email protected]
Spíchal Lukás
Lomin Sergey N
Strnad Miroslav
Schmülling Thomas
Article Info
Journal
Analytical biochemistry
Abbr.
Anal Biochem
ISSN
0003-2697
Published
2005-12-01
Epub
2005-00-30
Pages
129-34
Language
English
Region
United States
NLM ID
0370535
Subset
IM
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