Dataset Information

A proteome-wide map of 20(S)-hydroxycholesterol interactors in cell membranes

ABSTRACT: Oxysterols (OHCs) are hydroxylated cholesterol metabolites that play ubiquitous roles in health and disease. Due to the non-covalent nature of their interactions and unique partitioning in membranes, the analysis of live-cell, proteome-wide interactions of OHCs remains an unmet challenge. In this Resource, we present a structurally precise chemoproteomics probe for the osteogenic molecule 20(S)-hydroxycholesterol (20(S)-OHC) and provide a map of its proteome-wide targets in the membranes of living cells. Our target catalogue consolidates diverse OHC ontologies and demonstrates that OHC-interacting proteins cluster with specific processes in immune response and cancer. Competition experiments reveal that 20(S)-OHC is a chemo-, regio-, and stereoselective ligand for the protein Tmem97 (σ2 receptor), enabling molecular reconstruction of the Tmem97:20(S)-OHC binding site. Our results demonstrate that multiplexed, quantitative analysis of cellular target engagement can expose new dimensions of OHC activity and identify actionable targets for molecular therapy.

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture, Fibroblast

SUBMITTER: Brett Lomenick

LAB HEAD: Alison E. Ondrus

PROVIDER: PXD027787 | Pride | 2021-11-29

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	Yu-Shiuan_13Mar20_DMSO-1.raw.raw	Raw
	Yu-Shiuan_13Mar20_DMSO-2.raw	Raw
	Yu-Shiuan_13Mar20_Sequest_Reviewed.msf	Msf
	Yu-Shiuan_13Mar20_XM-1.raw	Raw
	Yu-Shiuan_13Mar20_XM-2.raw	Raw

Items per page:

1 - 5 of 10

Publications

A proteome-wide map of 20(S)-hydroxycholesterol interactors in cell membranes.

Cheng Yu-Shiuan YS Zhang Tianyi T Ma Xiang X Pratuangtham Sarida S Zhang Grace C GC Ondrus Alexander A AA Mafi Amirhossein A Lomenick Brett B Jones Jeffrey J JJ Ondrus Alison E AE

Nature chemical biology 20211119 12

Oxysterols (OHCs) are hydroxylated cholesterol metabolites that play ubiquitous roles in health and disease. Due to the non-covalent nature of their interactions and their unique partitioning in membranes, the analysis of live-cell, proteome-wide interactions of OHCs remains an unmet challenge. Here, we present a structurally precise chemoproteomics probe for the biologically active molecule 20(S)-hydroxycholesterol (20(S)-OHC) and provide a map of its proteome-wide targets in the membranes of l ...[more]

PMID: 34799735

Similar Datasets

Project description:Chemoselective modification of peptides and proteins has wide applications in chemical biology and pharmaceutical development. A highly efficient chemo-, regio- and stereoselective hydrosulfuration of ynamide was developed by taking advantage of an unprecedented β-addition of the thiols. Importantly, this reaction has been identified as an effective tool for Cys modification of peptides and proteins. The rationally designed strong electron-withdrawing triflyl group on the nitrogen atom of ynamides played a crucial role for controlling the chemo-, regio- and stereoselectivities while another substituent offered a handle for functionality diversification. This Cys modification strategy proceeded efficiently in a slightly basic aqueous conditions (pH 8) to provide exclusively the Z-isomer of the corresponding conjugates with superior stability. All the reactive peptide side chain functional groups such as amino, carboxyl, primary amide, and hydroxyl groups, as well as the unprotected imidazole and indole NH are compatible. This method displayed a broad substrate scope including linear and cyclic peptides, proteins and antibody. The potential application of this method in peptide and protein chemical biology was further exemplified by Cys-bioconjugation with ynamides containing functional molecules including small molecule drugs, fluorescent and affinity tags. In addition, this strategy was also compatible with click chemistry (performed in one-pot), which remarkably extended the application of this tool. Furthermore, the chemoselective biotinylation of ubiquitin(G47C) variant with a biotinylated ynamide, as well as the regioselective modification of Cys14 and Cys38 in bovine pancreatic trypsin inhibitor (BPTI), Cys34 of BSA and the antibody (Trastuzumab), could be accomplished readily under the optimized reaction conditions without perturbation of the other disulfide bonds. This method offered a novel and robust platform for Cys modification and opened new horizons for the production of peptide/protein/antibody-drug conjugates.

Project description:Hydroxylated polychlorinated biphenyls are the metabolites produced from polychlorinated biphenyls (PCBs) by drug-metabolizing enzyme cytochrome P450 1A1. These compounds are bound to transthyretin, a major plasma thyroid hormone-binding protein in amphibian tadpoles. The compounds-transthyretin complexes are transferred into the brain across the blood brain barrier in mammals. Thus these compounds are suspected to disrupt neural development in brain. We studied about the effects of hydroxylated PCBs on the thyroid system in brain using metamorphosing tadpoles of African clawed toad, Xenopus laevis. The metamorphosis assay revealed that these compounds had inhibitory effects on the thyroid hormone-induced metamorphosis. This in vivo assay was a powerful tool to detect thyroid-disrupting activities, because we were not able to detect the inhibitory effects of these compounds using thyroid hormone-responsive reporter gene assay in a cultured Xenopus cell line. A genome-wide gene expression analysis in brain following short-term exposure to these compounds demonstrated that the delay of metamorphosis and the morphological thyroid-disrupting changes could be caused partially by disruption of the thyroid hormone-induced gene expression by hydroxylated PCBs. Furthermore, we associated functional ontology terms with the transcripts whose expression were altered by thyroid hormone alone, or thyroid hormone and hydroxylated PCBs. We suggested that these approachs using a technique of bioinformatics revealed molecular mechanism of thyroid-disrupting activities in vivo. Thyroid hormones induce amphibian metamorphosis and alter a lot of thyroid hormone-responsive gene expression. We studied about the effects of hydroxylated PCBs on TH-induced gene expression. Premetamorphic tadpoles were treated with 500 nM hydroxylated PCBs in the presence of 1 nM thyroid hormone for 4 days. After exposure period total RNA was extracted from brain. Study included at least three replicate of each treatment.