化学
荧光
全内反射荧光显微镜
连接器
部分
生物物理学
立体化学
膜
生物化学
计算机科学
量子力学
生物
操作系统
物理
作者
Christoph Müller,Jakob Gleixner,Maris‐Johanna Tahk,Sergei Kopanchuk,Tõnis Laasfeld,Michael A. K. Weinhart,Dieter Schollmeyer,Martin U. Betschart,Steffen Lüdeke,Pierre Koch,Ago Rinken,Max Keller
标识
DOI:10.1021/acs.jmedchem.1c02033
摘要
The recent crystallization of the neuropeptide Y Y1 receptor (Y1R) in complex with the argininamide-type Y1R selective antagonist UR-MK299 (2) opened up a new approach toward structure-based design of nonpeptidic Y1R ligands. We designed novel fluorescent probes showing excellent Y1R selectivity and, in contrast to previously described fluorescent Y1R ligands, considerably higher (∼100-fold) binding affinity. This was achieved through the attachment of different fluorescent dyes to the diphenylacetyl moiety in 2 via an amine-functionalized linker. The fluorescent ligands exhibited picomolar Y1R binding affinities (pKi values of 9.36-9.95) and proved to be Y1R antagonists, as validated in a Fura-2 calcium assay. The versatile applicability of the probes as tool compounds was demonstrated by flow cytometry- and fluorescence anisotropy-based Y1R binding studies (saturation and competition binding and association and dissociation kinetics) as well as by widefield and total internal reflection fluorescence (TIRF) microscopy of live tumor cells, revealing that fluorescence was mainly localized at the plasma membrane.
科研通智能强力驱动
Strongly Powered by AbleSci AI