The role of cluster formation and metastable liquid - Liquid phase separation in protein crystallization

@article{Zhang2012TheRO,
  title={The role of cluster formation and metastable liquid - Liquid phase separation in protein crystallization},
  author={Fajun Zhang and Felix Roosen‐Runge and Andrea Sauter and Roland Roth and Maximilian W. A. Skoda and Robert M. J. Jacobs and Michael Sztucki and Frank Schreiber},
  journal={Faraday Discussions},
  year={2012},
  volume={159},
  pages={313-325}
}
We discuss the phase behavior and in particular crystallization of a model globular protein (beta-lactoglobulin) in solution in the presence of multivalent electrolytes. It has been shown previously that negatively charged globular proteins at neutral pH in the presence of multivalent counterions undergo a “re-entrant condensation (RC)” phase behavior (Zhang et al., Phys. Rev. Lett., 2008, 101, 148101), i.e. a phase-separated regime occurs in between two critical salt concentrations, c* < c… 

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References

SHOWING 1-10 OF 54 REFERENCES
Charge-controlled metastable liquid-liquid phase separation in protein solutions as a universal pathway towards crystallization
We demonstrate that a metastable liquid–liquid phase separation (LLPS) in protein aqueous solutions can be induced by multivalent metal ions at room temperature. We determine the salt and protein
Control of protein crystal nucleation around the metastable liquid-liquid phase boundary.
  • O. Galkin, P. Vekilov
  • Materials Science
    Proceedings of the National Academy of Sciences of the United States of America
  • 2000
TLDR
The rate of homogeneous nucleation of lysozyme crystals passes through a maximum in the vicinity of the liquid-liquid phase boundary hidden below the liquidus (solubility) line in the phase diagram of the protein solution, found that glycerol and polyethylene glycol shift this phase boundary and significantly suppress or enhance the crystal nucleation rates.
Toward Further Understanding of Lysozyme Crystallization: Phase Diagram, Protein−Protein Interaction, Nucleation Kinetics, and Growth Kinetics
A detailed study on the phase behavior and mechanism of lysozyme crystallization is presented. The nucleation and crystal growth rates, crystal morphologies, solubility, second virial coefficients,
Equilibrium cluster formation in concentrated protein solutions and colloids
TLDR
It is demonstrated that in both model systems, a combination of short-range attraction and long-range repulsion results in the formation of small equilibrium clusters, which is relevant for nucleation processes during protein crystallization, protein or DNA self-assembly.
Dense Liquid Precursor for the Nucleation of Ordered Solid Phases from Solution, Crystal Growth and Design
A line of recent theories and simulations have suggested that the nucleation of protein crystals might, under certain conditions, proceed in two steps:  the formation of a droplet of a dense liquid,
Liquid-Liquid Phase Separation in Supersaturated Lysozyme Solutions and Associated Precipitate Formation/Crystallization
Using cloud point determinations, the phase boundaries (binodals) for metastable liquid–liquid (L–L) separation in supersaturated hen egg white lysozyme solutions with 3%, 5%, and 7% (w/v) NaCl at
The two-step mechanism of nucleation of crystals in solution.
TLDR
This mechanism helps to explain several long-standing puzzles of crystal nucleation in solution: nucleation rates which are many orders of magnitude lower than theoretical predictions, nucleation kinetic dependencies with steady or receding parts at increasing supersaturation, the role of heterogeneous substrates for polymorph selection, the significance of the dense protein liquid, and others.
Quantitative imaging by confocal scanning fluorescence microscopy of protein crystallization via liquid-liquid phase separation.
TLDR
The results demonstrate the substantial possible complexity of crystallization behavior, as well as its sensitivity to the location of the conditions on the phase diagram and to the physicochemical properties of the system.
Metastable liquid clusters in super- and undersaturated protein solutions.
TLDR
A simple Monte Carlo model with protein-like potentials reproduces the metastable clusters of dense liquid with limited lifetimes and variable sizes and suggests that the mean cluster size is determined by the kinetics of growth and decay and not by thermodynamics.
Novel approach to controlled protein crystallization through ligandation of yttrium cations
Crystal structure determination of macromolecules is often hampered by the lack of crystals suitable for diffraction experiments. This article describes a protocol to crystallize the acidic protein
...
1
2
3
4
5
...