Arabidopsis HY5 protein functions as a DNA-binding tag for purification and functional immobilization of proteins on agarose/DNA microplate

FEBS Lett. 2008 Jan 23;582(2):221-8. doi: 10.1016/j.febslet.2007.12.004. Epub 2007 Dec 17.


Protein microarray is considered to be one of the key analytical tools for high-throughput protein function analysis. Here, we report that the Arabidopsis HY5 functions as a novel DNA-binding tag (DBtag) for proteins. We also demonstrate that the DBtagged proteins could be immobilized and purified on a newly designed agarose/DNA microplate. Furthermore, we show three applications using the microarray: (1) detection of autophosphorylation activity of DBtagged human protein kinases and inhibition of their activity by staurosporine, (2) specific cleavage of DBtagged proteins by a virus protease and caspase 3, and (3) detection of a protein-protein interaction between the DBtagged UBE2N and UBE2v1. Thus, this method may facilitate rapid functional analysis of a wide range of proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / physiology*
  • Arabidopsis Proteins / physiology*
  • Base Sequence
  • Basic-Leucine Zipper Transcription Factors / physiology*
  • DNA Primers
  • DNA-Binding Proteins / isolation & purification
  • DNA-Binding Proteins / physiology*
  • Nuclear Proteins / physiology*
  • Plant Proteins / isolation & purification
  • Plant Proteins / physiology*
  • Protein Array Analysis
  • Sepharose


  • Arabidopsis Proteins
  • Basic-Leucine Zipper Transcription Factors
  • DNA Primers
  • DNA-Binding Proteins
  • HY5 protein, Arabidopsis
  • Nuclear Proteins
  • Plant Proteins
  • Sepharose