Inhibition of OSBP blocks retrograde trafficking by inducing partial Golgi degradation

Endogenous and fluorescent sterols reveal the molecular basis for ligand selectivity of human sterol transporters

“…This result is consistent with the recently reported importance of an amide linker between the sterol core and sidechain, which is suggested to interact with Thr491, conferring tight OSBP binding and high selectivity. 17 Furthermore, TF-TMR-chol could serve as an alternative probe to the NBD-labeled probes for different purposes, since it is excited and emits at different wavelengths (Supplementary Figure 2). In contrast, TF-chol shows an overall weaker binding to the STPs suggesting that a longer linker between the cholesterol core and the BODIPY group might be preferred.…”

“…The screening platform, as such, enabled the identification of potent and selective OSBP binders, inhibiting retrograde trafficking and reducing Shiga toxin toxicity. 17 …”

“…The screening platform, as such, enabled the identification of potent and selective OSBP binders, inhibiting retrograde trafficking and reducing Shiga toxin toxicity. 17 Here we describe the profiling of fluorescent sterol-based probes as well as a set of steroidbased natural products, utilizing the now more comprehensive STP screening panel, providing the molecular basis for ligand recognition of ten STPs for the first time combined in one study. The identification of suitable fluorescent probes for the differential STPs enabled the establishment of fluorescence-, FRET-, and differential scanning fluorimetry-based assays, facilitating the direct comparison of STP specific selectivity profiles and evaluation of selective STP inhibitors in a high throughput manner.…”