Cosolvent Modulation of Protein Interactions especially for Protein Crystallization

Methylamine Osmolytes in Protein Crystallography

Osmolytes are organic solutes made by marine organisms for maintaining osmotic balance in saline environments. At Whitman, Biologist Paul Yancey investigates the constallation of osmlytes used by various marine organisms as a function of habitat (e.g. high vs low pressure). Osmolytes modulate both protein stability and solubility, and one osmolyte in particular, TMAO (trimethylamine N-oxide), is well-known for upstabilizing proteins. We are studying TMAO and other methylamine osmolytes as tools for crystallization and cryoprotection. We have found TMAO (trimethylamine N-oxide) TMAO (trimethylamine N-oxide to be an effective primary precipitant for protein crystallization. As well, TMAO, sarcosine, and betaine are all effective cryoprotective agents for cryocrystallography.


PEG Induced Crystallization

Polyethylene glycol (PEG) is a well-known polymer that sees multiple uses, including in the growth of macromolecular crystals. PEG induced macromolecular crystallization is thought to function via an entropic mechanism called the depletion effect. Here the PEG molecules, which are somewhat smaller than the protein molecules, are excluded from the protein surface because of their finite size. If the protein molecules associate, then the volume of solution accessible to the PEG actually increases, which is entropically favorable. If enough PEG molecules are in the solution, then this can actually drive the protein molecules together into a crystal. Recent solution studies looking at a liquid-liquid phase transition have suggested that small molecular weight PEGs may instead produce an effective repulsion between protein molecules. We are investigating whether this is true for protein crystallization.