A medium-throughput crystallization approach.
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.
Sulzenbacher
Gerlind
G
AFMB, UMR 6098, CNRS and Universités Aix-Marseille I and II, 31 Chemin J Aiguier, F-13402 Marseille CEDEX 20, France.
Gruez
Arnaud
A
Roig-Zamboni
Véronique
V
Spinelli
Silvia
S
Valencia
Christel
C
Pagot
Fabienne
F
Vincentelli
Renaud
R
Bignon
Christophe
C
Salomoni
Aurelia
A
Grisel
Sacha
S
Maurin
Damien
D
Huyghe
Céline
C
Johansson
Kent
K
Grassick
Alice
A
Roussel
Alain
A
Bourne
Yves
Y
Perrier
Sophie
S
Miallau
Linda
L
Cantau
Phillippe
P
Blanc
Eric
E
Genevois
Michel
M
Grossi
Alain
A
Zenatti
André
A
Campanacci
Valérie
V
Cambillau
Christian
C
D003460:Crystallization / Q000295:instrumentation / Q000379:methods*; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D008027:Light; D012371:Robotics; D012542:Scattering, Radiation
D029968:Escherichia coli Proteins