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

2021 ◽  
Author(s):  
Yu-Shiuan Cheng ◽  
Tianyi Zhang ◽  
Xiang Ma ◽  
Sarida Pratuangtham ◽  
Grace Zhang ◽  
...  
Keyword(s):  
Immune Response ◽  
Quantitative Analysis ◽  
Living Cells ◽  
Molecular Therapy ◽  
Interacting Proteins ◽  
Target Engagement ◽  
Cellular Target ◽  
Health And Disease ◽  
Cell Proteome ◽  
Covalent Nature

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.

Rapid Evaluation of Small Molecule Cellular Target Engagement with a Luminescent Thermal Shift Assay

2021 ◽  
Author(s):  
Jonathan D. Mortison ◽  
Ivan Cornella-Taracido ◽  
Gireedhar Venkatchalam ◽  
Anthony W. Partridge ◽  
Nirodhini Siriwardana ◽  
...  
Keyword(s):  
Small Molecule ◽  
Rapid Evaluation ◽  
Target Engagement ◽  
Thermal Shift Assay ◽  
Cellular Target ◽  
Thermal Shift

scholarly journals Evaluation of the binding affinity of E3 ubiquitin ligase ligands by cellular target engagement and in-cell ELISA assay

2021 ◽  
Vol 2 (1) ◽  
pp. 100288
Author(s):  
Ka Yang ◽  
Yaxian Zhou ◽  
Brett L. Roberts ◽  
Xueqing Nie ◽  
Weiping Tang
Keyword(s):  
Binding Affinity ◽  
Ubiquitin Ligase ◽  
E3 Ubiquitin Ligase ◽  
Elisa Assay ◽  
Target Engagement ◽  
Cellular Target ◽  
Cell Elisa

scholarly journals Small molecules, big effects: microbial metabolites in intestinal immunity

2020 ◽  
Vol 318 (5) ◽  
pp. G907-G911
Author(s):  
Lila G. Glotfelty ◽  
Andrea C. Wong ◽  
Maayan Levy
Keyword(s):  
Immune Response ◽  
Immune System ◽  
Microbial Community ◽  
Small Molecules ◽  
Defense Mechanisms ◽  
Microbial Metabolites ◽  
Intestinal Immunity ◽  
Vast Number ◽  
Health And Disease ◽  
Mammalian Intestine

The mammalian intestine is host to a vast number of microbial organisms. The immune system must balance tolerance with innate and adaptive defense mechanisms to maintain homeostasis with the microbial community. Interestingly, microbial metabolites have been shown to play a role in shaping the host immune response, thus assisting with adaptations that have significant implications for human health and disease. New investigations have uncovered roles for metabolites in modulating almost every aspect of the immune system. In this minireview, we survey these recent findings, which taken together reveal nuanced interactions that we are just beginning to understand.

Quantifying Target Occupancy of Small Molecules Within Living Cells

2020 ◽  
Vol 89 (1) ◽  
pp. 557-581 ◽  
Author(s):  
M.B. Robers ◽  
R. Friedman-Ohana ◽  
K.V.M. Huber ◽  
L. Kilpatrick ◽  
J.D. Vasta ◽  
...  
Keyword(s):  
Cultured Cells ◽  
Drug Efficacy ◽  
Living Cells ◽  
Drug Binding ◽  
Therapeutic Window ◽  
Target Engagement ◽  
Binding Properties ◽  
Structure Activity ◽  
Sar Analysis ◽  
Kinetics Of

The binding affinity and kinetics of target engagement are fundamental to establishing structure–activity relationships (SARs) for prospective therapeutic agents. Enhancing these binding parameters for operative targets, while minimizing binding to off-target sites, can translate to improved drug efficacy and a widened therapeutic window. Compound activity is typically assessed through modulation of an observed phenotype in cultured cells. Quantifying the corresponding binding properties under common cellular conditions can provide more meaningful interpretation of the cellular SAR analysis. Consequently, methods for assessing drug binding in living cells have advanced and are now integral to medicinal chemistry workflows. In this review, we survey key technological advancements that support quantitative assessments of target occupancy in cultured cells, emphasizing generalizable methodologies able to deliver analytical precision that heretofore required reductionist biochemical approaches.

Micropatterning for quantitative analysis of protein-protein interactions in living cells

2008 ◽  
Vol 5 (12) ◽  
pp. 1053-1060 ◽  
Author(s):  
Michaela Schwarzenbacher ◽  
Martin Kaltenbrunner ◽  
Mario Brameshuber ◽  
Clemens Hesch ◽  
Wolfgang Paster ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Protein Interactions ◽  
Living Cells ◽  
Protein Protein Interactions

Rapid discovery of drug target engagement by isothermal shift assay

2019 ◽  
Author(s):  
Kristofor J. Webb ◽  
Kerri A. Ball ◽  
Stephen J. Coleman ◽  
Jeremy Jacobsen ◽  
Michael H.B. Stowell ◽  
...  
Keyword(s):  
Drug Target ◽  
Drug Targets ◽  
Kinase Inhibitors ◽  
Rapid Identification ◽  
Living Cells ◽  
Experimental Designs ◽  
Living Systems ◽  
Target Engagement ◽  
Protein Targets ◽  
Drug Molecules

Identifying protein targets directly bound by drug molecules within living systems remains challenging. Here we present the isothermal shift assay, iTSA, for rapid identification of drug targets. Compared with thermal proteome profiling, a prevailing method for target engagement, iTSA offers a simplified workflow, 4-fold higher throughput, and multiplexed experimental designs with higher replication. We demonstrate application of iTSA to identify targets for several kinase inhibitors in lysates and living cells.

Rotation of the cell nucleus in living cells: a quantitative analysis

1985 ◽  
Vol 54 (2) ◽  
pp. 135-142 ◽  
Author(s):  
F. Bard ◽  
C. A. Bourgeois ◽  
D. Costagliola ◽  
M. Bouteille
Keyword(s):  
Quantitative Analysis ◽  
Cell Nucleus ◽  
Living Cells
Sign in / Sign up

Export Citation Format

Share Document