title: Characterization of gadolinium complexes for SAD phasing in macromolecular crystallography: application to CbpF.
abstract: Seven Gd complexes were used in the preparation of heavy-atom derivatives for solving the structure of choline-binding protein F (CbpF), a 36 kDa surface protein from Streptococcus pneumoniae, by the SAD method. CbpF was used as a model system to analyse the phasing capability of each of the derivatives. Three different aspects have been systematically characterized: the efficacy of cocrystallization versus soaking in the binding of the different Gd complexes, their mode of interaction and a comparative study of SAD phasing using synchrotron radiation and using a rotating-anode generator. This study reveals the striking potential of these complexes for SAD phasing using a laboratory source and further reinforces their relevance for high-throughput macromolecular crystallography.
authors.affiliation: Grupo de Cristalografía Macromolecular y Biología Estructural, Instituto Química Física Rocasolano, CSIC, Serrano 119, 28006 Madrid, Spain. xjuan@iqfr.csic.es
authors.forename: Rafael
authors.initials: R
authors.lastname: Molina
authors.forename: Meike
authors.initials: M
authors.lastname: Stelter
authors.forename: Richard
authors.initials: R
authors.lastname: Kahn
authors.forename: Juan A
authors.initials: JA
authors.lastname: Hermoso
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism; D002794:Choline / Q000378:metabolism; D003460:Crystallization; D005682:Gadolinium / Q000737:chemistry* / Q000378:metabolism; D046911:Macromolecular Substances / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D013296:Streptococcus pneumoniae; D013455:Sulfur / Q000737:chemistry / Q000378:metabolism; D017356:Synchrotrons; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D002352:Carrier Proteins; D046911:Macromolecular Substances; D013455:Sulfur; D005682:Gadolinium; D002794:Choline