Current technological background assigned for the crystallography of proteins in National Centre for Biomolecular Research

G. Demo¹, M. Wimmerova^1,2

¹National Centre for Biomolecular Research (NCBR), Kamenice 5, Brno, Czech Republic

²Department of Biochemistry, Kamenice 5, Brno, Czech Republic

guliver@mail.muni.cz

Keywords: glycobiochemistry group, technological background, lectins

The National Centre for Biomolecular Research (NCBR) is an independent unit at Masaryk University. NCBR is situated in the University Campus Bohunice, which is a complex of buildings used by Faculty of Medicine, Faculty of Science and Faculty of Sports. The NCBR is concerned with several areas of biological and chemical research, one of them is protein crystallography.

It is known that for the crystallography a crystal is needed. The devices used for the crystallisation available at NCBR, which allow an automatic screening of crystallisation experiments, are: Automatic liquid handling system Tecan Evo 150 and Crystallisation robot Mosquito. Tecan Evo 150 is used mainly for pipetting/mixing of crystallisation solutions into 96 well screening plates (reservoir). Mosquito is used for pipetting of a protein and crystallisation solutions in nanoliter volumes into 96 well screening plates (mainly a sitting drop).

For tracing and observing of a crystalline structure Rigaku Desktop Minstrel UV with the Gallery 160 crystal hotel and optical spectroscope Leica with CCD kamera are used. Rigaku machines consists of a plate hotel, where all plates are stored, and the imager is able to take pictures of every drop not just in visible area but also in UV area of light. This is a very helpful method how to distinguish between protein and inorganic salt crystals. The optical spectroscope with polarization filter is used for observing any inside defect of crystal.

With a co-operation of Biotechnological Department of ASCR the crystals are tested for observing any diffraction and all diffraction data are collected on a synchrotron in Grenoble (ESRF).

During the talk, several examples from the field of carbohydrate binding proteins will be given [1-5]:

1. Pokorná, M.; Cioci, G.; Perret, S.; Rebuffet, E.; Kostlánová, N.; Adam, J.; Gilboa-Garber, N.; Mitchell, E.P.; Imberty, A.; Wimmerová, M.; Biochemistry 45 (24), (2006), 7501 - 7510

2. Kostlanova, N.; Mitchell E.P.; Lortat-Jacob H.; Oscarson S.; Lahmann M.; Gilboa-Garber N.; Chambat G.; Wimmerova M.; Imberty A.; Journal of Biological Chemistry 280, (2005), 27839 – 27849

3. Wimmerova M.; Mitchell E.; Sanchez J.F.; Gautier, C.; Imberty A.; Journal of Biological Chemistry 278, (2003), 27059 – 27067

4. Šulák, O.; Cioci, G.; Delia, M.; Lahnmann, M.; Varrot, A.; Imberty, A.; Wimmerová, M.; Structure 18, (2010), 59 - 72

5. Lameignere, E.; Malinovská, L.; Sláviková, M.; Duchaud, E.; Mitchell, E.P.; Varrot, A.; Šedo, O.; Imberty, A.; Wimmerová, M.; Biochemical Journal, 411, (2008), 307 - 318