Title: Photo-affinity and Metabolic Labeling Probes Based on the Opioid Alkaloids

The opioids are powerful analgesics yet possess contingencies that can lead to opioid-use disorder. Chemical probes derived from the opioid alkaloids can provide deeper insight into the molecular interactions in a cellular context. Here, we designed and developed photo-click morphine (PCM-2) as a photo-affinity probe based on morphine and dialkynyl-acetyl morphine (DAAM) as a metabolic acetate reporter based on heroin. Application of these probes to SH-SY5Y, HEK293T, and U2OS cells revealed that PCM-2 and DAAM primarily localize to the lysosome amongst other locations inside the cell by confocal microscopy and chemical proteomics. Interaction site identification by mass spectrometry revealed the mitochondrial phosphate carrier protein, solute carrier family 25 member 3, SLC25A3, and histone H2B as acylation targets of DAAM. These data illustrate the utility of chemical probes to measure localization and protein interactions in a cellular context and will inform the design of probes based on the opioids in the future. Leon Duque, Vallavoju, and colleagues report the design and development of photo-affinity and metabolic labeling probes based on the opioid alkaloids. These chemical probes are used to measure localization and protein interactions in a cellular context by confocal microscopy and chemical proteomics. These insights will inform the development of future probes inspired by the opioid alkaloids.+++ image