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A Calcium Bioluminescence Assay for Functional Analysis of Mosquito (Aedes aegypti) and Tick (Rhipicephalus microplus) G Protein-coupled Receptors
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Chapters
- 00:05Title
- 01:40Establishment of Stable Cell Lines
- 03:37Selection of Clonal Cell Lines
- 05:40Plate Preparation for the Calcium Bioluminescence Assay
- 07:20Calcium Bioluminescence Assay: Instrument Operation and Data Analysis
- 08:45Results: Calcium Bioluminescence Assay for Functional Analysis of G Protein-Coupled Receptors
- 10:22Conclusion
This protocol provides instructions for clonal-cell line selection and a calcium bioluminescence assay to analyze the structure-activity relationships of synthesized arthropod neuropeptides on their cognate GPCRs. This assay can be used for receptor deorphanization and structure-activity relationship studies for synthetic analog design and peptide/drug-lead discovery.
Tags
Calcium Bioluminescence AssayFunctional AnalysisMosquitoAedes AegyptiTickRhipicephalus MicroplusG Protein-coupled ReceptorsArthropod Hormone ReceptorsPesticidesKinin ReceptorsNeuropeptidesStructure-activity RelationshipsHeterologous Kinin Receptor-expressing SystemsBiostable Kinin AnalogsDiuretic ProcessesMyotropic ProcessesDigestive ProcessesSouthern Cattle TickBoophilus MicroplusAedes AegyptiCHO-K1 Cell LineCalcium LevelsPeptide ApplicationRecombinant CellsAequorin Protein
