High-molecular-weight polymers for protein crystallization: poly-gamma-glutamic acid-based precipitants.
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.
Hu
Ting Chou
TC
Department of Chemistry, University of York, York YO10 5YW, England.
Korczyńska
Justyna
J
Smith
David K
DK
Brzozowski
Andrzej Marek
AM
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
D011506:Proteins; C511775:poly(gamma-glutamic acid); D011099:Polyglutamic Acid; D014995:Xylosidases; D009113:Muramidase; D019747:Aldose-Ketose Isomerases; C023298:xylose isomerase