Design, Synthesis, and Evaluation of a Luminescent Cholesterol Mimic

The development of new chemical tools with improved properties is essential to chemical and cell biology. Of particular interest is the development of mimics of small molecules with important cellular function that allow the direct observation of their trafficking in a cell. To this end, a novel 15-azasterol has been designed and synthesized as a luminescent cholesterol mimic for the monitoring of cholesterol trafficking. The brightness of this probe, which is similar to 32-times greater than the widely used dehydroergosterol probe, is combined with resistance to photobleaching in solution and in human fibroblasts and an exceptionally large Stokes-like shift of similar to 150-200 nm. The photophysical properties of the probe have been studied experimentally and computationally, suggesting an intersystem crossing to the triplet excited state with subsequent phosphorescent decay. Molecular dynamics simulations show a similar binding mode of cholesterol and the azasterol probe to NPC proteins, demonstrating the structural similarity of the probe to cholesterol.

Automated summary

The novel 15-azasterol has been designed and synthesized as a luminescent cholesterol mimic for monitoring cholesterol trafficking. It exhibits a brightness similar to 32-times greater than widely used probes, along with resistance to photobleaching and a large Stokes-like shift of approximately 150-200 nm. Experimental and computational studies have revealed the photophysical properties of the probe, indicating an intersystem crossing to the triplet excited state with subsequent phosphorescent decay. Additionally, molecular dynamics simulations demonstrate the structural similarity of the probe to cholesterol in its binding mode to NPC proteins.