title: Arylfluorosulfates Inactivate Intracellular Lipid Binding Protein(s) through Chemoselective SuFEx Reaction with a Binding Site Tyr Residue.
abstract: Arylfluorosulfates have appeared only rarely in the literature and have not been explored as probes for covalent conjugation to proteins, possibly because they were assumed to possess high reactivity, as with other sulfur(VI) halides. However, we find that arylfluorosulfates become reactive only under certain circumstances, e.g., when fluoride displacement by a nucleophile is facilitated. Herein, we explore the reactivity of structurally simple arylfluorosulfates toward the proteome of human cells. We demonstrate that the protein reactivity of arylfluorosulfates is lower than that of the corresponding aryl sulfonyl fluorides, which are better characterized with regard to proteome reactivity. We discovered that simple hydrophobic arylfluorosulfates selectively react with a few members of the intracellular lipid binding protein (iLBP) family. A central function of iLBPs is to deliver small-molecule ligands to nuclear hormone receptors. Arylfluorosulfate probe 1 reacts with a conserved tyrosine residue in the ligand-binding site of a subset of iLBPs. Arylfluorosulfate probes 3 and 4, featuring a biphenyl core, very selectively and efficiently modify cellular retinoic acid binding protein 2 (CRABP2), both in vitro and in living cells. The X-ray crystal structure of the CRABP2-4 conjugate, when considered together with binding site mutagenesis experiments, provides insight into how CRABP2 might activate arylfluorosulfates toward site-specific reaction. Treatment of breast cancer cells with probe 4 attenuates nuclear hormone receptor activity mediated by retinoic acid, an endogenous client lipid of CRABP2. Our findings demonstrate that arylfluorosulfates can selectively target single iLBPs, making them useful for understanding iLBP function.
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Wentao
authors.initials: W
authors.lastname: Chen
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Jiajia
authors.initials: J
authors.lastname: Dong
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Lars
authors.initials: L
authors.lastname: Plate
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: David E
authors.initials: DE
authors.lastname: Mortenson
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Gabriel J
authors.initials: GJ
authors.lastname: Brighty
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Suhua
authors.initials: S
authors.lastname: Li
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Yu
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Andrea
authors.initials: A
authors.lastname: Galmozzi
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Peter S
authors.initials: PS
authors.lastname: Lee
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Jonathan J
authors.initials: JJ
authors.lastname: Hulce
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Benjamin F
authors.initials: BF
authors.lastname: Cravatt
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Enrique
authors.initials: E
authors.lastname: Saez
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Evan T
authors.initials: ET
authors.lastname: Powers
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Ian A
authors.initials: IA
authors.lastname: Wilson
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: K Barry
authors.initials: KB
authors.lastname: Sharpless
authors.affiliation: Department of Chemistry and §Department of Molecular and Experimental Medicine, ∥Department of Chemical Physiology, ⊥Department of Integrative Structural and Computational Biology, #The Skaggs Institute for Chemical Biology, The Scripps Research Institute , La Jolla, California 92037, United States.
authors.forename: Jeffery W
authors.initials: JW
authors.lastname: Kelly
mesh_terms: D001665:Binding Sites; D018929:Cell Culture Techniques; D018360:Crystallography, X-Ray; D050556:Fatty Acid-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D005461:Fluorine; D006367:HeLa Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008024:Ligands; D061986:MCF-7 Cells; D018168:Receptors, Retinoic Acid / Q000737:chemistry / Q000378:metabolism*; D013464:Sulfuric Acids / Q000737:chemistry / Q000378:metabolism*; D014443:Tyrosine / Q000737:chemistry*
chemical_list: D050556:Fatty Acid-Binding Proteins; D008024:Ligands; D018168:Receptors, Retinoic Acid; D013464:Sulfuric Acids; C082928:retinoic acid binding protein II, cellular; D005461:Fluorine; D014443:Tyrosine