Development of an automated large-scale protein-crystallization and monitoring system for high-throughput protein-structure analyses.
Protein crystallization remains one of the bottlenecks in crystallographic analysis of macromolecules. An automated large-scale protein-crystallization system named PXS has been developed consisting of the following subsystems, which proceed in parallel under unified control software: dispensing precipitants and protein solutions, sealing crystallization plates, carrying robot, incubators, observation system and image-storage server. A sitting-drop crystallization plate specialized for PXS has also been designed and developed. PXS can set up 7680 drops for vapour diffusion per hour, which includes time for replenishing supplies such as disposable tips and crystallization plates. Images of the crystallization drops are automatically recorded according to a preprogrammed schedule and can be viewed by users remotely using web-based browser software. A number of protein crystals were successfully produced and several protein structures could be determined directly from crystals grown by PXS. In other cases, X-ray quality crystals were obtained by further optimization by manual screening based on the conditions found by PXS.
Hiraki
Masahiko
M
Structural Biology Research Center, Photon Factory, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK), Japan.
Kato
Ryuichi
R
Nagai
Minoru
M
Satoh
Tadashi
T
Hirano
Satoshi
S
Ihara
Kentaro
K
Kudo
Norio
N
Nagae
Masamichi
M
Kobayashi
Masanori
M
Inoue
Michio
M
Uejima
Tamami
T
Oda
Shunichiro
S
Chavas
Leonard M G
LM
Akutsu
Masato
M
Yamada
Yusuke
Y
Kawasaki
Masato
M
Matsugaki
Naohiro
N
Igarashi
Noriyuki
N
Suzuki
Mamoru
M
Wakatsuki
Soichi
S
D001331:Automation; D019295:Computational Biology / Q000379:methods*; D003201:Computers; D003460:Crystallization / Q000379:methods*; D030562:Databases, Protein; D020407:Internet; D008956:Models, Chemical; D008958:Models, Molecular; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D012984:Software; D013329:Structure-Activity Relationship; D014584:User-Computer Interface
D011506:Proteins