Crystallization still represents the bottleneck in the process of protein structure determination at high resolution, despite high-throughput structural genomics programs require optimized crystallization strategies regarding crystal quality, time, success rate, reproducibility and used protein amount. On the other side, the development of suitable materials for controlled heterogeneous nucleation might facilitate biomacromolecular crystallization in a variety of experimental conditions which are not conventionally fruitful. Here we show the possibility to fabricate hydrogel membranes displaying controlled chemical composition and nanostructure and to use them as heterogeneous supports for biomacromolecular crystallization. Diverse gel morphologies were obtained by controlling phase separation kinetics during gel layer formation on membrane support. These composite materials were found to increase the efficiency of the crystallization process so that crystals with enhanced diffraction properties were produced at lower protein concentration than conventional technique, thus affording the possibility to improve current approaches to protein crystallization and to be adapted to specific targets.

Tailored hydrogel membranes for efficient protein crystallization

Di Profio G;Belviso;Caliandro R;Fontananova E;Curcio E;Drioli;
2014

Abstract

Crystallization still represents the bottleneck in the process of protein structure determination at high resolution, despite high-throughput structural genomics programs require optimized crystallization strategies regarding crystal quality, time, success rate, reproducibility and used protein amount. On the other side, the development of suitable materials for controlled heterogeneous nucleation might facilitate biomacromolecular crystallization in a variety of experimental conditions which are not conventionally fruitful. Here we show the possibility to fabricate hydrogel membranes displaying controlled chemical composition and nanostructure and to use them as heterogeneous supports for biomacromolecular crystallization. Diverse gel morphologies were obtained by controlling phase separation kinetics during gel layer formation on membrane support. These composite materials were found to increase the efficiency of the crystallization process so that crystals with enhanced diffraction properties were produced at lower protein concentration than conventional technique, thus affording the possibility to improve current approaches to protein crystallization and to be adapted to specific targets.
2014
Istituto di Cristallografia - IC
Istituto per la Tecnologia delle Membrane - ITM
gels
composite membranes
protein crystallization
X-ray diffraction
crystal engineering
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.14243/262281
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