ИСТИНА

Two novel 20-hydroxy-pregn-5-en-3b-ols with NBD-label in their side-chains, attached to C20 via -NH2-CH2CH2- (1) and -NH2-CH2CH2CC- (2) linkers, were synthesized aiming to create fluorescent probes with 20-hydroxycholesterol-like scaffold. We estimated their interactions with whole cells of yeasts Yarrowia lipolytica and Saccharomyces cerevisiae at aerobic conditions. Using confocal microscopy the compounds were found to stain different compartments of the yeasts, including intracellular. Using chromatography and mass-spectrometry the steroids were also found to undergo partial conversion into 3-O-acetates by S. cerevisiae, but not Y. lipolytica, which was in accordance with computational BLAST search of S. cerevisiae Atf2p homologues. Also the compounds’ binding with mammalian STARD1 protein were observed. Compound 2 is also promising as a Raman probe due to alkyne insertion into its side chain. Basing on both the results and Autodock Vina-aided docking simulations with various sterol-binding proteins, e.g. STARD1 (3pl0), Osh4 (1zhw), Osh1 (5wvr) and Npc1 (3gkj), we speculate about possibilities of 20-hydroxycholesterol-like compounds to affect yeast cells or mammalian mitochondria as well as future perspectives of such compounds synthesis and usage for studies of 20-oxysterol trafficking and signaling. This work was supported by joint BRFFI (X16P-065) - RFFI (16-54-00139) grant, Belarusian SPSI grant 20161380 and RSF grant 17-74-10053.