title: A chemical proteomics approach for the search of pharmacological targets of the antimalarial clinical candidate albitiazolium in Plasmodium falciparum using photocrosslinking and click chemistry.
abstract: Plasmodium falciparum is responsible for severe malaria which is one of the most prevalent and deadly infectious diseases in the world. The antimalarial therapeutic arsenal is hampered by the onset of resistance to all known pharmacological classes of compounds, so new drugs with novel mechanisms of action are critically needed. Albitiazolium is a clinical antimalarial candidate from a series of choline analogs designed to inhibit plasmodial phospholipid metabolism. Here we developed an original chemical proteomic approach to identify parasite proteins targeted by albitiazolium during their native interaction in living parasites. We designed a bifunctional albitiazolium-derived compound (photoactivable and clickable) to covalently crosslink drug-interacting parasite proteins in situ followed by their isolation via click chemistry reactions. Mass spectrometry analysis of drug-interacting proteins and subsequent clustering on gene ontology terms revealed parasite proteins involved in lipid metabolic activities and, interestingly, also in lipid binding, transport, and vesicular transport functions. In accordance with this, the albitiazolium-derivative was localized in the endoplasmic reticulum and trans-Golgi network of P. falciparum. Importantly, during competitive assays with albitiazolium, the binding of choline/ethanolamine phosphotransferase (the enzyme involved in the last step of phosphatidylcholine synthesis) was substantially displaced, thus confirming the efficiency of this strategy for searching albitiazolium targets.
authors.affiliation: Dynamique des Interactions Membranaires Normales et Pathologiques, CNRS UMR 5235, Université Montpellier II, cc107, Place Eugène Bataillon, 34095 Montpellier Cedex 05, France.
authors.forename: Diana Marcela
authors.initials: DM
authors.lastname: Penarete-Vargas
authors.affiliation: Dynamique des Interactions Membranaires Normales et Pathologiques, CNRS UMR 5235, Université Montpellier II, cc107, Place Eugène Bataillon, 34095 Montpellier Cedex 05, France.
authors.forename: Anaïs
authors.initials: A
authors.lastname: Boisson
authors.affiliation: Institut de Génomique Fonctionnelle, CNRS UMR 5203, INSERM U661, Université Montpellier I, Université Montpellier II, F-34094 Montpellier, France.
authors.forename: Serge
authors.initials: S
authors.lastname: Urbach
authors.affiliation: Institut des Biomolécules Max Mousseron, CNRS UMR 5247, Université Montpellier II, cc1705, Place Eugène Bataillon, 34095 Montpellier Cedex 05, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Chantelauze
authors.affiliation: Institut des Biomolécules Max Mousseron, CNRS UMR 5247, Université Montpellier II, cc1705, Place Eugène Bataillon, 34095 Montpellier Cedex 05, France.
authors.forename: Suzanne
authors.initials: S
authors.lastname: Peyrottes
authors.affiliation: Sanofi, Therapeutic Strategic Unit for Infectious Diseases, 195 route d'Espagne, BP 13669, 31036 Toulouse Cedex, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Fraisse
authors.affiliation: Dynamique des Interactions Membranaires Normales et Pathologiques, CNRS UMR 5235, Université Montpellier II, cc107, Place Eugène Bataillon, 34095 Montpellier Cedex 05, France.
authors.forename: Henri J
authors.initials: HJ
authors.lastname: Vial
mesh_terms: D000818:Animals; D000962:Antimalarials / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D001667:Binding, Competitive; D057930:Click Chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry; D002798:Diacylglycerol Cholinephosphotransferase / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006801:Humans; D016778:Malaria, Falciparum / Q000378:metabolism / Q000469:parasitology / Q000517:prevention & control*; D008956:Models, Chemical; D015394:Molecular Structure; D010963:Plasmodium falciparum / Q000187:drug effects* / Q000378:metabolism; D011485:Protein Binding; D020543:Proteome / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism*; D013844:Thiazoles / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D021601:trans-Golgi Network / Q000378:metabolism
chemical_list: C499114:1,12-bis(4-methyl-5-(2-hydroxyethyl)thiazol-3-ium-3-yl)dodecane; D000962:Antimalarials; D003432:Cross-Linking Reagents; D020543:Proteome; D015800:Protozoan Proteins; D013844:Thiazoles; D002798:Diacylglycerol Cholinephosphotransferase