Work

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

Year: 2007

Type: article

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.

Source: FEBS Letters

DOI: 10.1016/j.febslet.2007.12.004

Authors: Tatsuya Sawasaki, Nami Kamura, Satoko Matsunaga, Mihoro Saeki, Masateru Tsuchimochi, Ryo Morishita, Yaeta Endo

Institutions: Ehime University, CellFree Sciences (Japan), RIKEN

Language: English

Topic: Advanced Biosensing Techniques and Applications

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: bronze

Funder: Ministry of Education, Culture, Sports, Science and Technology

OpenAlex ID: W2013833085

FWCI: 1.306

Cites: 32

Cited by: 67

Related to: 10