PUBLICATION
Combining Neurobehavioral Analysis and In Vivo Photoaffinity Labeling to Understand Protein Targets of Methamphetamine in Casper Zebrafish
- Authors
- Tackie-Yarboi, E., Wisner, A., Horton, A., Chau, T.Q.T., Reigle, J., Funk, A.J., Mccullumsmith, R.E., Hall, F.S., Williams, F.E., Schiefer, I.T.
- ID
- ZDB-PUB-200814-8
- Date
- 2020
- Source
- ACS Chemical Neuroscience 11(17): 2761-2773 (Journal)
- Registered Authors
- Keywords
- none
- MeSH Terms
-
- Animals
- Methamphetamine*
- Photoaffinity Labels
- Proteins
- Proteomics
- Zebrafish*
- PubMed
- 32786314 Full text @ ACS Chem. Neurosci.
Citation
Tackie-Yarboi, E., Wisner, A., Horton, A., Chau, T.Q.T., Reigle, J., Funk, A.J., Mccullumsmith, R.E., Hall, F.S., Williams, F.E., Schiefer, I.T. (2020) Combining Neurobehavioral Analysis and In Vivo Photoaffinity Labeling to Understand Protein Targets of Methamphetamine in Casper Zebrafish. ACS Chemical Neuroscience. 11(17):2761-2773.
Abstract
Photoaffinity labeling remains one of the most widely utilized methods of determining protein targets of drugs. Although useful, the scope of this technique has been limited to in vitro applications due to the inability of UV light to penetrate whole organisms. Herein, pigment-free Casper zebrafish were employed to allow in vivo photoaffinity labeling. A methamphetamine related phenethylamine photoaffinity labeling probe, designated here as 2, demonstrated dose-dependent effects on behavior similar to methamphetamine and permitted concentration dependent labeling of protein binding partners. Click chemistry was used to analyze binding partners via fluorimaging. Conjugation to a biotin permitted streptavidin pull-down and proteomic analysis to define direct binding partners of the methamphetamine probe. Bioinformatic analysis revealed the probe was chiefly bound to proteins involved in phagocytosis and mitochondrial function. Future applications of this experimental paradigm combining examination of drug-protein binding interactions alongside neurobehavioral readouts via in vivo PAL will significantly enhance our understanding of drug targets, mechanism(s) of action, and toxicity/lethality.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping