Protein crystallization occurs when the concentration of protein in solution is greater that its limit of solubility and so the protein is in a supersaturated zone Try to arrange Precipitation conditions such Nucleation 8 Clear that crystals (metastable will start to grow from saturated Crystallizing agent concentration protein solution Supersaturation Undersaturation
Protein crystallization occurs when the concentration of protein in solution is greater that its limit of solubility and so the protein is in a supersaturated zone Try to arrange conditions such that crystals will start to grow from saturated protein solution
rotein Hanging gla plate solution dr ro seal a watched crystal precipitant” mother never grows liquor rystallographer's Water will evaporate from protein drop fortune cookie to make precipitant concentration equal to that in reservoir, slowly ta king the protein into the crystallization region
Hanging drop Water will evaporate from protein drop to make precipitant concentration equal to that in reservoir, slowly taking the protein into the crystallization region ”mother liquor” A watched crystal never grows Crystallographer's fortune cookie
影响蛋白质结晶的因素 Physical Chemical Biochemical temperature pH value Purity of macromolecule surfaces precipitant type Ligands inhibitors effectors approach to equilibrium precipitant concentration Aggregation state gravity ionic strength Posttranslational modification pressure specitic ions Source of macromolecule time degree of supersaturation Proteolysis/hydrolysis vibrations reductive oxidative env. Chemical modifications electrostatic fields marcormolecule conc Genetic modifications magnetic fields metal ions Inherent symmetry of macromolecule dielectric of medium crosslinkers Stability of macromolecule viscosity of medium detergents/surfactants Isoelectric point rate of equilibration impurities Histrory of the sample homogeneous or heterogeneous nucleants Protein crystallization: more art than science!
影响蛋白质结晶的因素 Physical Chemical Biochemical temperature pH value Purity of macromolecule surfaces precipitant type Ligands, inhibitors, effectors approach to equilibrium precipitant concentration Aggregation state gravity ionic strength Posttranslational modification pressure specific ions Source of macromolecule time degree of supersaturation Proteolysis/hydrolysis vibrations reductive/oxidative env. Chemical modifications electrostatic fields marcormolecule conc. Genetic modifications magnetic fields metal ions Inherent symmetry of macromolecule dielectric of medium crosslinkers Stability of macromolecule viscosity of medium detergents/surfactants Isoelectric point rate of equilibration impurities Histrory of the sample homogeneous or heterogeneous nucleants Protein crystallization: more art than science!
Glutamate dehydrogenase F.Pennovorans pullulanase Glutaredoxin 3 S cerevisiae lumazine synthase Isocitrate dehydrogenase Rieske protein Gelatinase A-MMP2
F.Pennovorans pullulanase S.cerevisiae lumazine synthase Rieske protein Glutamate dehydrogenase Gelatinase A – MMP2 Isocitrate dehydrogenase Glutaredoxin 3
结晶机器人:高通量晶体生长 99999999 多(3(3(
结晶机器人：高通量晶体生长