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Fall 2017 Colloquium: Professor Allie Obermeyer
Tuesday, October 17, 2017, 10:30am - 11:30pm
 

obermeyerProfessor Allie Obermeyer

Columbia University

Tuesday October 17, 2017

Wright-Reiman Labs Room 260

"Insights into liquid-liquid demixing of globular proteins"

Globular proteins can undergo a liquid-liquid phase separation, termed complex coacervation, with oppositely charged polyelectrolytes. This macromolecule rich liquid phase can be used to stabilize the protein component, mimic the cytoplasmic environment, or enhance protein activity. However, due to the relatively low charge density and “patchy” charged surface of most proteins, many globular proteins do not phase separate with oppositely charged polymers. Efforts to understand the parameters that govern globular protein complex coacervation have focused on commercially available proteins and chemically modified variants of these proteins. These studies have revealed the importance of the ratio of charged residues on the protein and localized “charge patches.” To develop design criteria and engineering strategies for globular protein complex coacervation, additional understanding of the factors that control protein-polyelectrolyte phase separation is needed.

Genetic modification of recombinantly expressed proteins enables the systematic study of protein parameters that influence complex coacervation. Using green fluorescent protein (GFP) as a model protein, the impact of charge anisotropy on globular protein phase separation was investigated. Supercharged variants of GFP were biosynthesized with random distribution of charge on the protein surface. The net charge of the protein was varied from +30 to -30. Concurrently, the wild type GFP (with a net charge of -7) was modified at the C-terminus with a polyionic amino acid tag to vary the overall charge from +30 to -30 and provide an anisotropic, localized charged patch. Comparison of isotropic and anisotropic charge distribution on GFP provides insight into the mechanism by which charge localization impacts complex coacervation in these systems.

~Coffee/tea will be served prior to lecture~

 

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