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Title
Japanese: 
English:Immobilization of proteins onto microbeads using a DNA binding tag for enzymatic assays. 
Author
Japanese: Takaaki Kojima, Takuro Mizoguchi, Eri Ota, Jumpei Hata, Keisuke Homma, 朱 博, Kiyotaka Hitomi, Hideo Nakano.  
English: Takaaki Kojima, Takuro Mizoguchi, Eri Ota, Jumpei Hata, Keisuke Homma, Bo Zhu, Kiyotaka Hitomi, Hideo Nakano.  
Language English 
Journal/Book name
Japanese:Journal of bioscience and bioengineering 
English:Journal of bioscience and bioengineering 
Volume, Number, Page        
Published date Oct. 27, 2015 
Publisher
Japanese: 
English: 
Conference name
Japanese: 
English: 
Conference site
Japanese: 
English: 
Official URL https://doi.org/10.1016/j.jbiosc.2015.06.003
 
DOI https://doi.org/10.1016/j.jbiosc.2015.06.003
Abstract A novel DNA-binding protein tag, scCro-tag, which is a single-chain derivative of the bacteriophage lambda Cro repressor, has been developed to immobilize proteins of interest (POI) on a solid support through binding OR consensus DNA (ORC) that is tightly bound by the scCro protein. The scCro-tag successfully bound a transglutaminase 2 (TGase 2) substrate and manganese peroxidase (MnP) to microbeads via scaffolding DNA. The resulting protein-coated microbeads can be utilized for functional analysis of the enzymatic activity using flow cytometry. The quantity of bead-bound proteins can be enhanced by increasing the number of ORCs. In addition, proteins with the scCro-tag that were synthesized using a cell-free protein synthesis system were also immobilized onto the beads, thus indicating that this bead-based system would be applicable to high-throughput analysis of various enzymatic activities.

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