A novel approach for affinity-based screening of target specific ligands: Application of photoreactive D-glyceraldehyde-3-phosphate dehydrogenase
A novel approach for affinity-based screening of target specific ligands: Application of photoreactive D-glyceraldehyde-3-phosphate dehydrogenase
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DOI：
10.1021/bc0340520
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发表时间：
2003-09-01
影响因子：
4.7
通讯作者：
Hatanaka, Y
中科院分区：
文献类型：
--
作者：
Kaneda, M;Sadakane, Y;Hatanaka, Y
光亲和技术的新应用已被开发用于有效发现小配体和大分子相互作用。该方法称为光亲和捕获，使用光反应蛋白和固定配体来快速筛选竞争性抑制剂。制备了光反应性3-磷酸甘油醛脱氢酶（photo-GAPDH）和固定化染料配体组，并作为模型系统进行了检查。 photo-GAPDH 被证明可以有效捕获固定的配体。当系统中包含非固定化竞争性配体时，根据配体的亲和力阻止捕获。本方法将为基于亲和力的配体库筛选提供有效的工具。
A novel application of the photoaffinity technique has been developed for the efficient discovery of small ligand and macromolecule interaction. The approach, photoaffinity capture, uses a photoreactive protein together with immobilized ligand for the rapid screening of competitive inhibitors. The set of photoreactive glyceraldehyde-3-phosphate dehydrogenase (photo-GAPDH) and immobilized dye ligand was prepared and examined as a model system. The photo-GAPDH was shown to efficiently capture the immobilized ligand. When nonimmobilized competitive ligands were included in the system, the capture was prevented in accordance with the affinity of the ligands. The present approach would provide an efficient tool for affinity-based screening of ligand libraries.