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Protein Crystallography

Description
Protein Crystallography activities of IMBB are equipped with appropriate incubators for bacterial cell growth, a fermentor, sonicators for cell lysis, protein purification apparatus (centrifuges, liquid chromatography systems, 2 FPLC systems), instrumentation for biophysical characterization of proteins (Circular Dichroism spectrometer, Mass spectrometry), controlled temperature room for crystallizations, equipment for X-ray diffraction data collection, structure determination and analysis employing suitable powerful computing systems.

Equipment
- Fermentor of 30 liter capacity (Bioengineering)

- ÄKTA FPLC (GE Healthcare-Pharmacia)

- CD-spectropolarimeter J-810 (Jasco)

- Room at constant temperature for crystallization of biomolecules.
For pictures see also:
Protein crystallography 1Protein Crystallography 2

- CAD4 X-ray diffractometer (Enraf-Nonius BV)

- X-ray rotating anode generator (RU-3HR, Rigaku), crystal cryo-cooling system (X-stream, Rigaku) and system of 'imaging-plate' photon detector (Mar300, MarResearch).
For pictures see also X-ray_generator 1, X-ray_generator 2

- Mar300 imaging plate X-ray detector

- Computing infrastructure for diffraction data processing, structure determination and analysis.
The above as well as the wet laboratories for protein purification and biophysical characterization are located on the ground floor of Biology Department next to a state of the art electron microscope facility of the University of Crete.

Services
For questions regarding to possible structure determination or modeling of a structure of a biomolecule, please refer to any of the following contact persons.

Contact Persons

Name Dr. Renate Gessmann
Telephone number +30-2810-394352
Fax Number +30-2810-394408
Email address This email address is being protected from spambots. You need JavaScript enabled to view it.
Room Number E128

 

Name Prof. Michael Kokkinidis
Telephone number +30-2810-394351
Fax Number +30-2810-394351
Email address This email address is being protected from spambots. You need JavaScript enabled to view it.
Room Number G124ß

 

Name Dr. Kyriacos Petratos
Telephone number +30-2810-394353
Fax Number +30-2810-394353
Email address This email address is being protected from spambots. You need JavaScript enabled to view it.
Room Number G124γ

 

Links (publications, manuals, analysis software)
- Fermentor
- ÄKTA FPLC
- CD-spectropolarimeter J-810
- CAD4 single crystal diffractometer
- Rigaku X-ray generator
- MarResearch imaging plate detector


Publications (that resulted from the use of the facility)
G. Prag, Y. Papanikolau, G. Tavlas, C. E. Vorgias, K. Petratos and A. B. Oppenheim (2000) Structures of chitobiase mutants complexed with the substrate di-N-acetyl-D-glucosamine: the catalytic role of the conserved acidic pair, aspartate 539 and glutamate 540 J. Mol. Biol. 300, 611-617.  

Y. Papanikolau, G. Prag, G. Tavlas, C. E. Vorgias, A. B. Oppenheim and K. Petratos (2001) High Resolution Structural Analyses of Mutant Chitinase A Complexes with Substrates Provide New Insight into the Mechanism of Catalysis Biochemistry 40, 11338-11343.

Y. Papanikola u and K. Petratos (2002) Application of the effects of ionic strength reducing agents in the purification and crystallization of chitinase A Acta Crystallogr. D 58, 1593-1596.

Y. Papanikolau, G. Tavlas, C. E. Vorgias and K. Petratos (2003) De novo purification scheme and crystallization conditions yield high-resolution structures of chitinase A and its complex with the inhibitor allosamidin Acta Crystallogr. D 59, 400-403.

R. Gessmann, H. Brückner and K. Petratos (2003) Three Complete Turns of a 3 10 -Helix at Atomic Resolution: The Crystal Structure of Z-(Aib) 11 -O t Bu J. Peptide Sci. 9, 753-762.

Y. Papanikolau , I . Tsigos , M . Papadovasilaki , V . Bouriotis and K . Petratos (2005) Crystallization and preliminary X - ray diffraction studies of an alcohol dehydrogenase from the Antarctic psychrophile Moraxella sp . TAE 123 Acta Crystallogr. F 61, 246-248.

Y. Papanikolau, M. Papadovasilaki, R. B. G. Ravelli, A. A. McCarthy, S. Cusack , A. Economou and K. Petratos (2007) Structure of dimeric SecA , the Escherichia coli preprotein translocase motor J. Mol. Biol. 366, 1545-1557.

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