Selective inhibition of OSBP blocks retrograde trafficking by inducing partial Golgi degradation

Abstract

Sterol-binding proteins are important regulators of lipid homeostasis and membrane integrity; however, the discovery of selective small molecule modulators can be challenging due to structural similarities in the sterol binding domains. We report the discovery of highly potent and selective inhibitors of oxysterol binding protein (OSBP), which we term oxybipins. Sterol-containing chemical chimeras aimed at identifying new sterol binding proteins by targeted degradation, led to a significant reduction in Golgi-associated proteins. The degradation was found to occur at lysosomes, concomitant with changes in general protein glycosylation, indicating that the degradation of Golgi proteins was a downstream effect. By establishing a sterol transport protein biophysical assay panel, we discovered that the oxybipins potently inhibited OSBP, resulting in blockage of retrograde trafficking and attenuating Shiga toxin toxicity. As the oxybipins do not target any other sterol transporters tested, we advocate their use as chemical tools to study OSBP function and therapeutic relevance.

Orpinolide disrupts a leukemic dependency on cholesterol transport by inhibiting OSBP

“…In vitro fluorescence polarization assays with a recombinant glutathione S-transferase (GST)-tagged sterol-binding domain of OSBP (OSBP-related domain or ORD) 37 indicated that orpinolide engages OSBP in the nanomolar range and that binding is selective over other sterol transporters 38,39 (Fig. 4e and Extended Data Fig.…”