title: One assay for all: exploring small molecule phosphorylation using amylose-polyiodide complexes.
abstract: We present a generic method for screening small molecule kinases for their acceptor specificity. The release of the reaction byproduct adenosine diphosphate (ADP) triggers a concentration-dependent formation of amylose from sucrose, by using the combined enzymatic action of sucrose synthase and glycogen synthase. Kinase activities could be quantified photometrically after the formation of a dark-blue amylose-polyiodide complex. We demonstrate that this method can be used to profile both known and novel nucleotide- and sugar-kinases for their substrate specificity. Using a facile and widely available methodology, the amylose-polyiodide small-molecule kinase assay presented herein has the potential to perform substrate screenings of small molecule kinases in a high-throughput manner.
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University , Nanjing, Jiangsu 210095 China.
authors: Xu C
authors: XC
authors: Duan
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University , Nanjing, Jiangsu 210095 China.
authors: Huan
authors: H
authors: Chen
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University , Nanjing, Jiangsu 210095 China.
authors: Fang F
authors: FF
authors: Liu
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University , Nanjing, Jiangsu 210095 China.
authors: Louis
authors: L
authors: Conway
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University , Nanjing, Jiangsu 210095 China.
authors: Shuang
authors: S
authors: Wei
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University , Nanjing, Jiangsu 210095 China.
authors: Zhi P
authors: ZP
authors: Cai
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University , Nanjing, Jiangsu 210095 China.
authors: Li
authors: L
authors: Liu
authors: Glycomics and Glycan Bioengineering Research Center (GGBRC), College of Food Science and Technology, Nanjing Agricultural University , Nanjing, Jiangsu 210095 China.
authors: Josef
authors: J
authors: Voglmeir
mesh_terms: D000688:Amylose / Q000737:chemistry* / Q000378:metabolism; D003124:Colorimetry; D007455:Iodine / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation; D010770:Phosphotransferases / Q000032:analysis* / Q000378:metabolism
chemical_list: D000688:Amylose; D007455:Iodine; D010770:Phosphotransferases