About: 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.   Goto Sponge  NotDistinct  Permalink

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  • 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.
Subject
  • Virology
  • Proteomics
  • Arabidopsis
  • DNA-binding proteins
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