One Assay for All: Exploring Small Molecule Phosphorylation Using Amylose–Polyiodide Complexes
| Autor: | Fang F Liu, Louis P. Conway, Huan Chen, Josef Voglmeir, Li Liu, Zhi P. Cai, Xu C. Duan, Shuang Wei |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
chemistry.chemical_classification
biology Kinase Phosphotransferases Substrate (chemistry) Small molecule Analytical Chemistry Adenosine diphosphate chemistry.chemical_compound Enzyme chemistry Biochemistry biology.protein Sucrose synthase Phosphorylation Colorimetry Amylose Glycogen synthase Iodine |
| Zdroj: | Analytical Chemistry. 87:9546-9550 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.5b02247 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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