Unknown

Dataset Information

0

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.

SUBMITTER: Sawasaki T 

PROVIDER: S-EPMC7164004 | biostudies-literature | 2008 Jan

REPOSITORIES: biostudies-literature

altmetric image

Publications

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

Sawasaki Tatsuya T   Kamura Nami N   Matsunaga Satoko S   Saeki Mihoro M   Tsuchimochi Masateru M   Morishita Ryo R   Endo Yaeta Y  

FEBS letters 20071217 2


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  ...[more]

Similar Datasets

| S-EPMC2795123 | biostudies-literature
| S-EPMC3495177 | biostudies-literature
| S-EPMC427747 | biostudies-literature
| S-EPMC3175489 | biostudies-literature
| S-EPMC5500333 | biostudies-literature
| S-EPMC5645317 | biostudies-literature
| S-EPMC6128974 | biostudies-literature
| S-EPMC5491447 | biostudies-literature
| S-EPMC3406683 | biostudies-literature
| S-EPMC4853705 | biostudies-other