Adoff Hayden, Halls Victoria S, Holland Emily, Lobingier Braden, Arttamangkul Seksiri
Chemical Physiology and Biochemistry, School of Medicine, Oregon Health and Science University, Portland, OR, USA.
Medicinal Chemistry Core Facility, Oregon Health and Science University, Portland, OR, USA.
STAR Protoc. 2023 Apr 26;4(2):102231. doi: 10.1016/j.xpro.2023.102231.
This protocol describes endogenous labeling of opioid receptors (ORs) using a ligand-directed reagent, naltrexamine-acylimidazole compounds (NAI-X). NAI acts by guiding and permanently tagging a small-molecule reporter (X)-such as fluorophores or biotin-to ORs. Here we detail syntheses and uses of NAI-X for OR visualization and functional studies. The NAI-X compounds overcome long-standing challenges in mapping and tracking endogenous ORs as the labeling can be done in situ with live tissues or cultured cells. For complete details on the use and execution of this protocol, please refer to Arttamangkul et al..
本方案描述了使用配体导向试剂纳曲胺-酰基咪唑化合物（NAI-X）对内源性阿片受体（ORs）进行标记。NAI通过引导并将小分子报告基团（X）——如荧光团或生物素——永久标记到ORs上发挥作用。在这里，我们详细介绍了NAI-X的合成及其用于OR可视化和功能研究的方法。NAI-X化合物克服了在绘制和追踪内源性ORs方面长期存在的挑战，因为这种标记可以在活组织或培养细胞中原位进行。有关本方案使用和实施的完整详细信息，请参考Arttamangkul等人的文献。
