title: A medium-throughput crystallization approach.
abstract: The first results of a medium-scale structural genomics program clearly demonstrate the value of using a medium-throughput crystallization approach based on a two-step procedure: a large screening step employing robotics, followed by manual or automated optimization of the crystallization conditions. The structural genomics program was based on cloning in the Gateway vectors pDEST17, introducing a long 21-residue tail at the N-terminus. So far, this tail has not appeared to hamper crystallization. In ten months, 25 proteins were subjected to crystallization; 13 yielded crystals, of which ten led to usable data sets and five to structures. Furthermore, the results using a robot dispensing 50-200 nl drops indicate that smaller protein samples can be used for crystallization. These still partial results might indicate present and future directions for those who have to make crucial choices concerning their crystallization platform in structural genomics programs.
authors.affiliation: AFMB, UMR 6098, CNRS and Universités Aix-Marseille I and II, 31 Chemin J Aiguier, F-13402 Marseille CEDEX 20, France.
authors.forename: Gerlind
authors.initials: G
authors.lastname: Sulzenbacher
authors.forename: Arnaud
authors.initials: A
authors.lastname: Gruez
authors.forename: Véronique
authors.initials: V
authors.lastname: Roig-Zamboni
authors.forename: Silvia
authors.initials: S
authors.lastname: Spinelli
authors.forename: Christel
authors.initials: C
authors.lastname: Valencia
authors.forename: Fabienne
authors.initials: F
authors.lastname: Pagot
authors.forename: Renaud
authors.initials: R
authors.lastname: Vincentelli
authors.forename: Christophe
authors.initials: C
authors.lastname: Bignon
authors.forename: Aurelia
authors.initials: A
authors.lastname: Salomoni
authors.forename: Sacha
authors.initials: S
authors.lastname: Grisel
authors.forename: Damien
authors.initials: D
authors.lastname: Maurin
authors.forename: Céline
authors.initials: C
authors.lastname: Huyghe
authors.forename: Kent
authors.initials: K
authors.lastname: Johansson
authors.forename: Alice
authors.initials: A
authors.lastname: Grassick
authors.forename: Alain
authors.initials: A
authors.lastname: Roussel
authors.forename: Yves
authors.initials: Y
authors.lastname: Bourne
authors.forename: Sophie
authors.initials: S
authors.lastname: Perrier
authors.forename: Linda
authors.initials: L
authors.lastname: Miallau
authors.forename: Phillippe
authors.initials: P
authors.lastname: Cantau
authors.forename: Eric
authors.initials: E
authors.lastname: Blanc
authors.forename: Michel
authors.initials: M
authors.lastname: Genevois
authors.forename: Alain
authors.initials: A
authors.lastname: Grossi
authors.forename: André
authors.initials: A
authors.lastname: Zenatti
authors.forename: Valérie
authors.initials: V
authors.lastname: Campanacci
authors.forename: Christian
authors.initials: C
authors.lastname: Cambillau
mesh_terms: D003460:Crystallization / Q000295:instrumentation / Q000379:methods*; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D008027:Light; D012371:Robotics; D012542:Scattering, Radiation
chemical_list: D029968:Escherichia coli Proteins