title: Combining 'dry' co-crystallization and in situ diffraction to facilitate ligand screening by X-ray crystallography.
abstract: X-ray crystallography is an established technique for ligand screening in fragment-based drug-design projects, but the required manual handling steps - soaking crystals with ligand and the subsequent harvesting - are tedious and limit the throughput of the process. Here, an alternative approach is reported: crystallization plates are pre-coated with potential binders prior to protein crystallization and X-ray diffraction is performed directly 'in situ' (or in-plate). Its performance is demonstrated on distinct and relevant therapeutic targets currently being studied for ligand screening by X-ray crystallography using either a bending-magnet beamline or a rotating-anode generator. The possibility of using DMSO stock solutions of the ligands to be coated opens up a route to screening most chemical libraries.
authors.affiliation: CNRS, UMR5048 - Université de Montpellier, Centre de Biochimie Structurale, 34090 Montpellier, France.
authors.forename: Muriel
authors.initials: M
authors.lastname: Gelin
authors.affiliation: CNRS, UMR5048 - Université de Montpellier, Centre de Biochimie Structurale, 34090 Montpellier, France.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Delfosse
authors.affiliation: CNRS, UMR5048 - Université de Montpellier, Centre de Biochimie Structurale, 34090 Montpellier, France.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Allemand
authors.affiliation: CNRS, UMR5048 - Université de Montpellier, Centre de Biochimie Structurale, 34090 Montpellier, France.
authors.forename: François
authors.initials: F
authors.lastname: Hoh
authors.affiliation: Université Grenoble Alpes, IBS, 38044 Grenoble, France.
authors.forename: Yoann
authors.initials: Y
authors.lastname: Sallaz-Damaz
authors.affiliation: Université Grenoble Alpes, IBS, 38044 Grenoble, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Pirocchi
authors.affiliation: CNRS, UMR5048 - Université de Montpellier, Centre de Biochimie Structurale, 34090 Montpellier, France.
authors.forename: William
authors.initials: W
authors.lastname: Bourguet
authors.affiliation: Université Grenoble Alpes, IBS, 38044 Grenoble, France.
authors.forename: Jean Luc
authors.initials: JL
authors.lastname: Ferrer
authors.affiliation: CNRS, UMR5048 - Université de Montpellier, Centre de Biochimie Structurale, 34090 Montpellier, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Labesse
authors.affiliation: CNRS, UMR5048 - Université de Montpellier, Centre de Biochimie Structurale, 34090 Montpellier, France.
authors.forename: Jean François
authors.initials: JF
authors.lastname: Guichou
mesh_terms: D000818:Animals; D002645:Chickens; D003460:Crystallization / Q000379:methods*; D018360:Crystallography, X-Ray / Q000379:methods*; D000081406:Peptidyl-Prolyl Isomerase F; D021983:Cyclophilins / Q000737:chemistry / Q000378:metabolism; D055808:Drug Discovery / Q000379:methods*; D006801:Humans; D008024:Ligands; D019950:Mitogen-Activated Protein Kinase 1 / Q000737:chemistry / Q000378:metabolism; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism; D047495:PPAR gamma / Q000737:chemistry / Q000378:metabolism; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D051381:Rats
chemical_list: D000081406:Peptidyl-Prolyl Isomerase F; D008024:Ligands; D047495:PPAR gamma; D011506:Proteins; D019950:Mitogen-Activated Protein Kinase 1; C421864:hen egg lysozyme; D009113:Muramidase; D021983:Cyclophilins
keywords: drug design; fragment library screening; in-situ X-ray diffraction; ligand screening; therapeutic targets