Molecules (2021)
Update date:2022-07-29
Topics:
Boursier, Michelle E.
Hall, Mary P.
Hurst, Robin
Killoran, Michael P.
Kirkland, Thomas A.
Levin, Sergiy
Machleidt, Thomas
Ohana, Rachel Friedman
Zimmerman, Kristopher
Gaining insight into the pharmacology of ligand engagement with G-protein coupled receptors (GPCRs) under biologically relevant conditions is vital to both drug discovery and basic research. NanoLuc-based bioluminescence resonance energy transfer (NanoBRET) monitoring competitive binding between fluorescent tracers and unmodified test compounds has emerged as a robust and sensitive method to quantify ligand engagement with specific GPCRs genetically fused to NanoLuc luciferase or the luminogenic HiBiT peptide. However, development of fluorescent tracers is often challenging and remains the principal bottleneck for this approach. One way to alleviate the burden of developing a specific tracer for each receptor is using promiscuous tracers, which is made possible by the intrinsic specificity of BRET. Here, we devised an integrated tracer discovery workflow that couples machine learning-guided in silico screening for scaffolds displaying promiscuous binding to GPCRs with a blend of synthetic strategies to rapidly generate multiple tracer candidates. Subsequently, these candidates were evaluated for binding in a NanoBRET ligand-engagement screen across a library of HiBiT-tagged GPCRs. Employing this workflow, we generated several promiscuous fluorescent tracers that can effectively engage multiple GPCRs, demonstrating the efficiency of this approach. We believe that this workflow has the potential to accelerate discovery of NanoBRET fluorescent tracers for GPCRs and other target classes.
View MoreYancheng Smiling Imp & Exp Co., Ltd.
Contact:+86-515-83173586
Address:Rm1207, BLD#03, Phoenix Plaza, Juheng Road, Yancheng, Jiangsu, P.R. China
Chongqing Werlchem Fine Chemical Co. Ltd
Contact:+86-23-67521957
Address:NO.15,Fortune Road,Yubei District,401121,Chongqing,China
Zhejiang kehong chemical co., ltd
Contact:0086-575-85522000
Address:xiner center RD binhai industrial zone shaoxing zhejiang province P.R.China,312073
Hangzhou Ocean Chemical Co., Ltd.
website:http://www.hzoceanchem.com
Contact:+86-571-88025872, 28272092, 28272096
Address:Room 623 ,Building No 1 , COFCO Radius Commercial Center Xiwen Road, Xiacheng District, Hangzhou, Zhejiang Province, China
Contact:+1-973-357-0577
Address:10 Taft Rd.
Doi:10.1021/jo800822p
(2008)Doi:10.1021/jo00387a023
(1987)Doi:10.1021/ja00275a047
(1986)Doi:10.1002/chem.200801212
(2008)Doi:10.1007/s10593-008-0014-8
(2008)Doi:10.1248/cpb.34.1553
(1986)