The open-access high-throughput crystallization facility at EMBL Hamburg

Abstract

Here, the establishment of Europe's largest high-throughput crystallization facility with open access to the general user community is reported. The facility covers every step in the crystallization process from the preparation of crystallization cocktails for initial or customized screens to the setup of hanging-drop vapour-diffusion experiments and their automatic imaging. In its first year of operation, 43 internal and 40 external users submitted over 500 samples for a total of 2985 crystallization plates. An electronic booking system for registration, the selection of experimental parameters (e.g. drop size, sample-to-reservoir ratio) and the reservation of time slots was developed. External users can choose from more than 1000 initial crystallization conditions. By default, experiments are kept for six months and are imaged 15 times during this time period. A remote viewing system is available to inspect experiments via the internet. Over 100 stock solutions are available for users wishing to design customized screens.

Cloning, expression, purification and preliminary X-ray analysis of EstN2, a novel archaeal α/β-hydrolase fromCandidatusNitrososphaera gargensis

“…The final sample that was subjected to crystallization contained protein at a concentration of 10 mg ml À1 in a buffer consisting of 50 mM NaCl, 50 mM HEPES pH 7.5. The initial crystallization experiments were performed using commercial screens at the EMBL Hamburg high-throughput crystallization facility (Mueller-Dieckmann, 2006) and resulted in several good-quality crystals. The best data were collected from a crystal that diffracted to 1.5 Å resolution.…”

Structure-based Epitope Mapping of Mycobacterium tuberculosis Secretary Antigen MTC28

“…Screening of initial crystallization conditions for MTC28 was done by the sitting-drop vapor-diffusion method at 20 -25°C by mixing an equal volume of concentrated protein and reservoir solution. Initial crystallization trials were performed at the Indian Institute of Technology Kharagpur, India, and at the High-throughput Crystallization facility at European Molecular Biology Laboratory (EMBL), Hamburg (12). Of 735 tested conditions, 21 crystallization hits were identified.…”

Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of tetrahydrodipicolinate-N-succinyltransferase (Rv1201c) fromMycobacterium tuberculosis

“…Initial crystallization experiments were carried out at the High Throughput Crystallization Facility at the EMBL Hamburg Outstation (Mueller-Dieckmann, 2006). Initial crystals appeared within 4 d in Hampton Crystal Screen Lite condition No.…”