A new method of protein nucleation and crystallization based on Langmuir-Blodgett (LB) technology was utilized for the template stimulation of crystal growth of so far non-crystallized proteins. Microcrystals (60 - 120 micron) of bovine cytochrome P450scc and human protein kinase CKII alpha subunit were obtained with use of the homologous protein thin film template by vapor diffusion modified hanging drop method under all crystallization conditions including those failing to lead to crystal formation in solution. The induction of microcrystals nucleation by the thin template confirms in the two different important classes of proteins, until now never crystallized, the positive stimulatory influence for crystal formation of protein thin film template, which was observed in earlier study with a model system (lysozyme) as an unexpected acceleration and enhancement in the crystal growth. The presentation is focused on bovine cytochrome P450scc crystallization and subsequent crystals characterization by Atomic Force Microscopy under appropriate chamber. The results are discussed in terms of a possible transition in protein crystallization from art to science by means of thin film nanotechnology.
From art to science in protein crystallization by means of thin film technology – cytochrome P450scc crystals
PESHKOVA, EVGENIYA
2002-01-01
Abstract
A new method of protein nucleation and crystallization based on Langmuir-Blodgett (LB) technology was utilized for the template stimulation of crystal growth of so far non-crystallized proteins. Microcrystals (60 - 120 micron) of bovine cytochrome P450scc and human protein kinase CKII alpha subunit were obtained with use of the homologous protein thin film template by vapor diffusion modified hanging drop method under all crystallization conditions including those failing to lead to crystal formation in solution. The induction of microcrystals nucleation by the thin template confirms in the two different important classes of proteins, until now never crystallized, the positive stimulatory influence for crystal formation of protein thin film template, which was observed in earlier study with a model system (lysozyme) as an unexpected acceleration and enhancement in the crystal growth. The presentation is focused on bovine cytochrome P450scc crystallization and subsequent crystals characterization by Atomic Force Microscopy under appropriate chamber. The results are discussed in terms of a possible transition in protein crystallization from art to science by means of thin film nanotechnology.
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