title: Arabidopsis HY5 protein functions as a DNA-binding tag for purification and functional immobilization of proteins on agarose/DNA microplate.
abstract: 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.
authors: Cell-Free Science and Technology Research Center, and The Venture Business Laboratory, Ehime University, Matsuyama, Ehime, Japan. sawasaki@eng.ehime-u.ac.jp
authors: Tatsuya
authors: T
authors: Sawasaki
authors: Nami
authors: N
authors: Kamura
authors: Satoko
authors: S
authors: Matsunaga
authors: Mihoro
authors: M
authors: Saeki
authors: Masateru
authors: M
authors: Tsuchimochi
authors: Ryo
authors: R
authors: Morishita
authors: Yaeta
authors: Y
authors: Endo
mesh_terms: D017360:Arabidopsis / Q000502:physiology*; D029681:Arabidopsis Proteins / Q000502:physiology*; D001483:Base Sequence; D050976:Basic-Leucine Zipper Transcription Factors / Q000502:physiology*; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000302:isolation & purification / Q000502:physiology*; D009687:Nuclear Proteins / Q000502:physiology*; D010940:Plant Proteins / Q000302:isolation & purification / Q000502:physiology*; D040081:Protein Array Analysis; D012685:Sepharose
chemical_list: D029681:Arabidopsis Proteins; D050976:Basic-Leucine Zipper Transcription Factors; D017931:DNA Primers; D004268:DNA-Binding Proteins; C109254:HY5 protein, Arabidopsis; D009687:Nuclear Proteins; D010940:Plant Proteins; D012685:Sepharose