Protein-polymer interaction
Use of polymer-based precipitants is widely spread in the current protein crystallization screens. Molecules of poly-ethylene glycol has been shown to act as a “molecular glue” in formation of stable protein crystals in number of crystallization experiments. In spite of that, the variety of the used polymeric compounds remain limited. Therefore, variety of the protein - polymer interactions provided by commercial crystallization screens is also limited.
Our effort is focused on two following issues:
1) analysis of all available observed protein - polymer interactions
2) widening the spectrum of available polymer precipitants
Figure 1. Unusual case where the polymer molecule sticks together the neighbor protein molecules to form the crystal lattice. Six monomers of poly(ethyleneglycol) bind to asparagines of two neighbor protein molecules ensuring thus the specific adhesion decisive for the crystal packing shown here (PDB ID 1D1J).
Our analysis of the PDB records and of our own experiments has helped us to design two new polymer-based crystallization screens containing eight novel polymers that have not been tested before. We have successfully crystallized number of various proteins, some of them already known, and also several novel proteins like extracellular domain of mouse NKR-P1A and Organophosphorus acid anhydrolase from Alteromonas macleodii. We have also succeeded to collect the highest resolution diffraction data for the human protein CD69. Several other proteins crystallized using this screen are awaiting publication.
Figure 2. Crystals of xylanase in PolyB, no.41.
Papers published:
Skalova, T; Duskova, J; Hasek, J; Kolenko, P; Stepankova, A; Dohnalek, J.
Alternative polymer precipitants for protein crystallization
JOURNAL OF APPLIED CRYSTALLOGRAPHY, 2010, 43, 737-742.
Andrea Štěpánková, Jarmila Dušková, Tereza Skálová, Jindřich Hašek, Tomáš Koval', Lars Henrik Ostergaard and Jan Dohnálek
Organophosphorous acid anhydrolase from Alteromonas macleodii - a structural study and functional relationship to prolidases
Acta Cryst. (2013) F69, 346-354.
J. Hasek, J. Labsky, T. Skalova, P. Kolenko, J. Dohnalek, J. Duskova, A. Stepankova und T. Koval
Polymer structure database and protein-polymer interactions
Z. Kristallogr. Proc. 1 (2011) 475-480.
Kolenko, P; Skalova, T; Vanek, O; Stepankova, A; Duskova, J; Hasek, J; Bezouska, K; Dohnalek, J
The high-resolution structure of the extracellular domain of human CD69 using a novel polymer
ACTA CRYSTALLOGRAPHICA, 2009, F65, 1258-1260.
Hasek, J.
Poly(ethylene glycol) interactions with proteins
Z.Kristallogr.Suppl., 2006, 23, 613-618.