Probe-Based Chemical Proteomics Identifies UDP-Glucose 6-Dehydrogenase as a Potential Cannabidiol-Interacting Protein

Abstract

Cannabidiol (CBD) exhibits broad anti-inflammatory and neuroprotective effects. Although CBD has been reported to act on multiple molecular targets, the direct protein interactors and molecular mechanisms underlying its complex pharmacological activities remain incompletely understood. In this study, we developed a biotinylated CBD probe (II-2) and a FITC-labeled CBD fluorescent probe (III-3) for affinity enrichment and cellular imaging, respectively. Probe-mediated target fishing identified UDP-glucose dehydrogenase (UGDH) as a potential CBD-interacting protein. Competitive pull-down and western blot analysis showed that the CBD-derived probe specifically enriched UGDH, and this enrichment was competitively attenuated by excess CBD. Biolayer interferometry was consistent with an apparent micromolar affinity association between CBD and UGDH under the assay conditions, with an apparent K_d of approximately 5.8 μM. Functional assays showed that CBD enhanced UGDH enzymatic activity in a LPS-induced inflammatory model without markedly altering UGDH protein expression. Immunofluorescence imaging further supported the intracellular spatial association between the CBD-derived fluorescent probe and UGDH. These findings suggest that UGDH is a potential CBD-interacting protein and provide an experimental basis for further elucidating the molecular basis of CBD-related anti-inflammatory activity.