title: In Vitro Characterization of VPS34 Lipid Kinase Inhibition by Small Molecules.
abstract: VPS34 is a class III phosphoinositide 3-kinase that acts on vesicle trafficking. This kinase has recently attracted significant attention because of the function it plays in the machinery involved in the early steps of autophagy. Moreover, because significant progress had been made in the optimization of specific kinase inhibitors, its potential to be targeted by catalytic inhibitors has been investigated by different groups. The aim of this review is to present the key in vitro assays necessary for characterizing inhibitors of the catalytic activity of VPS34. The review covers catalytic (IC50on purified recombinant protein) and binding assays (KD, ka, kdon purified recombinant protein), and a cell-based assay (IC50in GFP-FYVE expressing cell line). The methodology for crystallization of VPS34 protein is also presented as it can provide guidance for the design by medicinal chemistry of small molecular mass kinase inhibitor.
authors.affiliation: Sanofi, Quai Jules Guesde, Vitry Sur Seine, Paris, France.
authors.forename: F
authors.initials: F
authors.lastname: Fassy
authors.affiliation: Sanofi, Quai Jules Guesde, Vitry Sur Seine, Paris, France.
authors.forename: C
authors.initials: C
authors.lastname: Dureuil
authors.affiliation: Sanofi, Quai Jules Guesde, Vitry Sur Seine, Paris, France.
authors.forename: A
authors.initials: A
authors.lastname: Lamberton
authors.affiliation: Sanofi, Quai Jules Guesde, Vitry Sur Seine, Paris, France.
authors.forename: M
authors.initials: M
authors.lastname: Mathieu
authors.affiliation: Sanofi, Quai Jules Guesde, Vitry Sur Seine, Paris, France.
authors.forename: N
authors.initials: N
authors.lastname: Michot
authors.affiliation: Sanofi, Quai Jules Guesde, Vitry Sur Seine, Paris, France.
authors.forename: B
authors.initials: B
authors.lastname: Ronan
authors.affiliation: Sanofi, Quai Jules Guesde, Vitry Sur Seine, Paris, France. Electronic address: benoit.pasquier@sanofi.com.
authors.forename: B
authors.initials: B
authors.lastname: Pasquier
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D001343:Autophagy; D058546:Class III Phosphatidylinositol 3-Kinases / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003460:Crystallization / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D000255:Adenosine Triphosphate; D058546:Class III Phosphatidylinositol 3-Kinases
keywords: Autophagy; Endosomes; Lipid kinase; PI3K; PIK3C3/VPS34; Vesicle trafficking