Endogenous and Fluorescent Sterols Reveal the Molecular Basis for Ligand Selectivity of Human Sterol Transporters

Overview

Journal J Lipid Res

Publisher Elsevier

Specialty Biochemistry

Date 2025 Jan 2

PMID 39746449

Authors

Laura Depta

Hogan P Bryce-Rogers

Nienke J Dekker

Anna Wiehl Bonke

Nicolo Camporese

Mingxing Qian

Yuanjian Xu

Douglas F Covey

Luca Laraia

Affiliations

Soon will be listed here.

Abstract

Sterol transport proteins (STPs) play a pivotal role in cholesterol homeostasis and therefore are essential for healthy human physiology. Despite recent advances in dissecting functions of STPs in the human cell, there is still a significant knowledge gap regarding their specific biological functions and a lack of suitable selective probes for their study. Here, we profile fluorescent steroid-based probes across ten STPs, uncovering substantial differences in their selectivity, aiding the retrospective and prospective interpretation of biological results generated with those probes. These results guided the establishment of an STP screening panel combining diverse biophysical assays, enabling the evaluation of 42 steroid-based natural products and derivatives. Combining this with a thorough structural analysis revealed the molecular basis for STP-specific selectivity profiles, leading to the uncovering of several new potent and selective Aster-B inhibitors and supporting the role of this protein in steroidogenesis.

Citing Articles

Identification of a Privileged Scaffold for Inhibition of Sterol Transport Proteins through the Synthesis and Ring Distortion of Diverse, Pseudo-Natural Products.

Bro F, Depta L, Dekker N, Bryce-Rogers H, Madsen M, Praestegaard K ACS Cent Sci. 2025; 11(1):136-146.

PMID: 39866705 PMC: 11758220. DOI: 10.1021/acscentsci.4c01657.

References

Luo J, Yang H, Song B . Mechanisms and regulation of cholesterol homeostasis. Nat Rev Mol Cell Biol. 2019; 21(4):225-245. DOI: 10.1038/s41580-019-0190-7. View

Lehto M, Laitinen S, Chinetti G, Johansson M, Ehnholm C, Staels B . The OSBP-related protein family in humans. J Lipid Res. 2001; 42(8):1203-13. View

Atkovska K, Klingler J, Oberwinkler J, Keller S, Hub J . Rationalizing Steroid Interactions with Lipid Membranes: Conformations, Partitioning, and Kinetics. ACS Cent Sci. 2018; 4(9):1155-1165. PMC: 6161064. DOI: 10.1021/acscentsci.8b00332. View

Storch J, Xu Z . Niemann-Pick C2 (NPC2) and intracellular cholesterol trafficking. Biochim Biophys Acta. 2009; 1791(7):671-8. PMC: 4281484. DOI: 10.1016/j.bbalip.2009.02.001. View

Tugaeva K, Faletrov Y, Allakhverdiev E, Shkumatov V, Maksimov E, Sluchanko N . Effect of the NBD-group position on interaction of fluorescently-labeled cholesterol analogues with human steroidogenic acute regulatory protein STARD1. Biochem Biophys Res Commun. 2018; 497(1):58-64. DOI: 10.1016/j.bbrc.2018.02.014. View

Iaea D, Gale S, Bielska A, Krishnan K, Fujiwara H, Jiang H . A novel intrinsically fluorescent probe for study of uptake and trafficking of 25-hydroxycholesterol. J Lipid Res. 2015; 56(12):2408-19. PMC: 4655980. DOI: 10.1194/jlr.D064287. View

Licciardello M, Workman P . The era of high-quality chemical probes. RSC Med Chem. 2022; 13(12):1446-1459. PMC: 9749956. DOI: 10.1039/d2md00291d. View

Selvaraj V, Stocco D, Clark B . Current knowledge on the acute regulation of steroidogenesis. Biol Reprod. 2018; 99(1):13-26. PMC: 6044331. DOI: 10.1093/biolre/ioy102. View

Akula N, Midzak A, Lecanu L, Papadopoulos V . Identification of small-molecule inhibitors of the steroidogenic acute regulatory protein (STARD1) by structure-based design. Bioorg Med Chem Lett. 2012; 22(12):4139-43. DOI: 10.1016/j.bmcl.2012.04.058. View

10.

Naito T, Ercan B, Krshnan L, Triebl A, Koh D, Wei F . Movement of accessible plasma membrane cholesterol by the GRAMD1 lipid transfer protein complex. Elife. 2019; 8. PMC: 6905856. DOI: 10.7554/eLife.51401. View

11.

He N, Depta L, Sievers S, Laraia L . Fluorescent probes and degraders of the sterol transport protein Aster-A. Bioorg Med Chem. 2024; 103:117673. DOI: 10.1016/j.bmc.2024.117673. View

12.

Rodriguez-Agudo D, Ren S, Wong E, Marques D, Redford K, Gil G . Intracellular cholesterol transporter StarD4 binds free cholesterol and increases cholesteryl ester formation. J Lipid Res. 2008; 49(7):1409-19. PMC: 2431108. DOI: 10.1194/jlr.M700537-JLR200. View

13.

Zhang J, Andersen J, Sun H, Liu X, Sonenberg N, Nie J . Aster-C coordinates with COP I vesicles to regulate lysosomal trafficking and activation of mTORC1. EMBO Rep. 2020; 21(9):e49898. PMC: 7507422. DOI: 10.15252/embr.201949898. View

14.

Wang H, Ma Q, Qi Y, Dong J, Du X, Rae J . ORP2 Delivers Cholesterol to the Plasma Membrane in Exchange for Phosphatidylinositol 4, 5-Bisphosphate (PI(4,5)P). Mol Cell. 2018; 73(3):458-473.e7. DOI: 10.1016/j.molcel.2018.11.014. View

15.

Sezgin E, Can F, Schneider F, Clausen M, Galiani S, Stanly T . A comparative study on fluorescent cholesterol analogs as versatile cellular reporters. J Lipid Res. 2015; 57(2):299-309. PMC: 4727425. DOI: 10.1194/jlr.M065326. View

16.

Szomek M, Moesgaard L, Reinholdt P, Haarhoj Hald S, Petersen D, Krishnan K . Membrane organization and intracellular transport of a fluorescent analogue of 27-hydroxycholesterol. Chem Phys Lipids. 2020; 233:105004. DOI: 10.1016/j.chemphyslip.2020.105004. View

17.

Wang P, Weng J, Lee S, Anderson R . The N terminus controls sterol binding while the C terminus regulates the scaffolding function of OSBP. J Biol Chem. 2008; 283(12):8034-45. DOI: 10.1074/jbc.M707631200. View

18.

Dong J, Du X, Wang H, Wang J, Lu C, Chen X . Allosteric enhancement of ORP1-mediated cholesterol transport by PI(4,5)P/PI(3,4)P. Nat Commun. 2019; 10(1):829. PMC: 6381110. DOI: 10.1038/s41467-019-08791-0. View

19.

Andersen J, Zhang J, Sun H, Liu X, Liu J, Nie J . Aster-B coordinates with Arf1 to regulate mitochondrial cholesterol transport. Mol Metab. 2020; 42:101055. PMC: 7476871. DOI: 10.1016/j.molmet.2020.101055. View

20.

Jaworski C, Moreira E, Li A, Lee R, Rodriguez I . A family of 12 human genes containing oxysterol-binding domains. Genomics. 2001; 78(3):185-96. DOI: 10.1006/geno.2001.6663. View