Combining Neurobehavioral Analysis and In Vivo Photoaffinity Labeling to Understand Protein Targets of Methamphetamine in Casper Zebrafish

• DOI: 10.1021/acschemneuro.0c00416
• Publication Date: 2020-07-31T01:10:55Z
• Authors: Ethel Tackie-Yarboi; Alexander Wisner; Austin Horton; Tue Q. T. Chau; James Reigle; Adam J. Funk; Robert E. McCullu...; Frank S. Hall; Frederick E. Will...; Isaac T. Schiefer
• Keywords: Photoaffinity labeling; Methamphetamine

Photoaffinity labeling (PAL) remains one of the most widely utilized methods determining protein targets drugs. Although useful, scope this technique has been limited to in vitro applications because inability UV light penetrate whole organisms. Herein, pigment-free Casper zebrafish were employed allow vivo PAL. A methamphetamine-related phenethylamine PAL probe, designated here as 2, demonstrated dose-dependent effects on behavior similar methamphetamine and permitted concentration-dependent binding partners. Click chemistry was used analyze partners via fluoroimaging. Conjugation a biotin streptavidin pull-down proteomic analysis define direct probe. Bioinformatic revealed probe chiefly bound proteins involved phagocytosis mitochondrial function. Future experimental paradigm combining examination drug–protein interactions alongside neurobehavioral readouts will significantly enhance our understanding drug targets, mechanism(s) action, toxicity/lethality.