title: Ligand and Target Discovery by Fragment-Based Screening in Human Cells.
abstract: Advances in the synthesis and screening of small-molecule libraries have accelerated the discovery of chemical probes for studying biological processes. Still, only a small fraction of the human proteome has chemical ligands. Here, we describe a platform that marries fragment-based ligand discovery with quantitative chemical proteomics to map thousands of reversible small molecule-protein interactions directly in human cells, many of which can be site-specifically determined. We show that fragment hits can be advanced to furnish selective ligands that affect the activity of proteins heretofore lacking chemical probes. We further combine fragment-based chemical proteomics with phenotypic screening to identify small molecules that promote adipocyte differentiation by engaging the poorly characterized membrane protein PGRMC2. Fragment-based screening in human cells thus provides an extensive proteome-wide map of protein ligandability and facilitates the coordinated discovery of bioactive small molecules and their molecular targets.
authors.affiliation: Department of Chemical Physiology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA. Electronic address: cparker@scripps.edu.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Parker
authors.affiliation: Department of Chemical Physiology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Andrea
authors.initials: A
authors.lastname: Galmozzi
authors.affiliation: Department of Chemical Physiology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Yujia
authors.initials: Y
authors.lastname: Wang
authors.affiliation: École Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland.
authors.forename: Bruno E
authors.initials: BE
authors.lastname: Correia
authors.affiliation: Department of Chemical Physiology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Kenji
authors.initials: K
authors.lastname: Sasaki
authors.affiliation: Department of Chemical Physiology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Joslyn
authors.affiliation: Department of Chemical Physiology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
authors.forename: Arthur S
authors.initials: AS
authors.lastname: Kim
authors.affiliation: Research and Development, Bristol-Myers Squibb Company, Princeton, NJ 08648, USA.
authors.forename: Cullen L
authors.initials: CL
authors.lastname: Cavallaro
authors.affiliation: Research and Development, Bristol-Myers Squibb Company, Princeton, NJ 08648, USA.
authors.forename: R Michael
authors.initials: RM
authors.lastname: Lawrence
authors.affiliation: Research and Development, Bristol-Myers Squibb Company, Princeton, NJ 08648, USA.
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Johnson
authors.affiliation: The Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Iñigo
authors.initials: I
authors.lastname: Narvaiza
authors.affiliation: Department of Chemical Physiology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA. Electronic address: esaez@scripps.edu.
authors.forename: Enrique
authors.initials: E
authors.lastname: Saez
authors.affiliation: Department of Chemical Physiology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, CA 92037, USA. Electronic address: cravatt@scripps.edu.
authors.forename: Benjamin F
authors.initials: BF
authors.lastname: Cravatt
mesh_terms: D017667:Adipocytes / Q000166:cytology; D002454:Cell Differentiation; D018360:Crystallography, X-Ray; D055808:Drug Discovery / Q000379:methods*; D057166:High-Throughput Screening Assays; D006801:Humans; D006867:Hydrolases / Q000737:chemistry; D008024:Ligands; D008565:Membrane Proteins / Q000037:antagonists & inhibitors; D010088:Oxidoreductases / Q000737:chemistry; D011485:Protein Binding; D040901:Proteomics / Q000379:methods*; D011980:Receptors, Progesterone / Q000037:antagonists & inhibitors; D054852:Small Molecule Libraries
chemical_list: D008024:Ligands; D008565:Membrane Proteins; C115297:PGRMC2 protein, human; D011980:Receptors, Progesterone; D054852:Small Molecule Libraries; D010088:Oxidoreductases; D006867:Hydrolases
keywords: FBLD; PGRMC2; adipogenesis; chemical probes; chemical proteomics; fragment-based ligand discovery; ligands; mass spectrometry; phenotypic screening; photoreactivity