title: High-molecular-weight polymers for protein crystallization: poly-gamma-glutamic acid-based precipitants.
abstract: Protein crystallization has been revolutionized by the introduction of high-throughput technologies, which have led to a speeding up of the process while simultaneously reducing the amount of protein sample necessary. Nonetheless, the chemistry dimension of protein crystallization has remained relatively undeveloped. Most crystallization screens are based on the same set of precipitants. To address this shortcoming, the development of new protein precipitants based on poly-gamma-glutamic acid (PGA) polymers with different molecular-weight ranges is reported here: PGA-LM (low molecular weight) of approximately 400 kDa and PGA-HM (high molecular weight) of >1,000 kDa. It is also demonstrated that protein precipitants can be expanded further to polymers with much higher molecular weight than those that are currently in use. Furthermore, the modification of PGA-like polymers by covalent attachments of glucosamine substantially improved their solubility without affecting their crystallization properties. Some preliminary PGA-based screens are presented here.
authors.affiliation: Department of Chemistry, University of York, York YO10 5YW, England.
authors.forename: Ting Chou
authors.initials: TC
authors.lastname: Hu
authors.forename: Justyna
authors.initials: J
authors.lastname: Korczyńska
authors.forename: David K
authors.initials: DK
authors.lastname: Smith
authors.forename: Andrzej Marek
authors.initials: AM
authors.lastname: Brzozowski
mesh_terms: D019747:Aldose-Ketose Isomerases / Q000737:chemistry; D003460:Crystallization / Q000379:methods*; D008970:Molecular Weight; D009113:Muramidase / Q000737:chemistry; D011099:Polyglutamic Acid / Q000031:analogs & derivatives* / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D014995:Xylosidases / Q000737:chemistry
chemical_list: D011506:Proteins; C511775:poly(gamma-glutamic acid); D011099:Polyglutamic Acid; D014995:Xylosidases; D009113:Muramidase; D019747:Aldose-Ketose Isomerases; C023298:xylose isomerase