Post-biotinylation of Photocrosslinking by Staudinger-Bertozzi Ligation of Preinstalled Alkylazide Tag
作者:Makoto Hashimoto、Yasumaru Hatanaka
DOI:10.1248/cpb.53.1510
日期:——
Post-biotinylation of the alkyl azide derivative of trifluoromethyl phenyldiazirine (TPD) was elucidated to apply a photoaffinity biotinylation technique. A photo-modified polyvinilidene difluoride (PVDF) membrane was used as a photolabeled component and we introduced biotin by Staudinger–Bertozzi ligation. The 15 pmol amount of biotinylated reagent was still effective for the visualization of cross-linked product on the matrix. The results show the potential utility of alkyl azide carrying TPD derivatives in the application of photoaffinity biotinylation, which could be useful for the ligands with tight structural requirements.
通过对三氟甲基苯二氮丙啶(TPD)的烷基叠氮化物衍生物进行生物素化后处理,阐明了光亲和生物素化技术的应用。我们使用光修饰的聚偏二氟乙烯(PVDF)膜作为光标记成分,并通过施陶丁格-贝托齐(Staudinger-Bertozzi)连接法引入生物素。15 pmol 量的生物素化试剂仍能有效地观察基质上的交联产物。结果表明,烷基叠氮化物携带的 TPD 衍生物在光亲和生物素化应用中具有潜在的实用性,可用于结构要求严格的配体。