Microdrop screening: a rapid method to optimize solvent conditions for NMR spectroscopy of proteins.
Determining appropriate solvent conditions is a crucial first step for carrying out NMR spectroscopy of proteins, but rapid and efficient methods for doing so are currently lacking. Microdrop screening examines a large number of different solvent conditions using very small amounts of protein and minimal labor. Starting from one initial buffer condition, small aliquots of protein solution are combined with an array of solutions in which concentration, pH, buffer type, and added stabilizers are systematically varied. The protein concentration of each microliter-sized test drop ('microdrop') is gradually changed using vapor diffusion, and the solubility of the protein is determined by visual examination. A variety of analytical techniques may be applied to the contents of the microdrops to monitor enzymatic activity, aggregation, ligand binding, and protein folding.
Lepre
C A
CA
Vertex Pharmaceuticals, Inc., Cambridge, MA 02139-4242, USA.
Moore
J M
JM
D002021:Buffers; D020904:Glia Maturation Factor; D006801:Humans; D009414:Nerve Growth Factors / Q000737:chemistry; D009419:Nerve Tissue Proteins / Q000737:chemistry*; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D012997:Solvents
D002021:Buffers; D020904:Glia Maturation Factor; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D011506:Proteins; D011994:Recombinant Proteins; D012997:Solvents