Single molecule imaging techniques to aid crystallization

摘要

This invention relates to a method to rapidly find crystallization conditions for a biomolecule in a desired conformation using single-molecule, fluorescent resonance energy transfer (smFRET) imaging techniques. The method provides significant cost and time advantages over the empricial exploration for crystallization conditions.

主权项

1. A method for selecting a crystallization for a biomolecule comprising:
(a) labeling the biomolecule with a donor and acceptor fluorophore sufficiently spaced to report time-dependent dynamics of multiple conformations of the biomolecule by single molecule fluorescence resonance energy transfer (smFRET), wherein said multiple conformations includes a first conformation; (b) placing the labeled biomolecule under different crystallization conditions; (c) detecting fluorescence resonance energy transfer between the donor and acceptor fluorophores for each condition over a period of time; (d) determining the amount of time the biomolecule spends in said first conformation, relative to other conformation or conformations; and (e) selecting the crystallization condition under which the biomolecule spends more time in said first conformation relative to other conformation or conformations.