Dynamic and Asymmetric Protein Microcompartmentation in Aqueous Two-Phase Vesicles
Open Access
Author:
Long, Michael Scott
Graduate Program:
Chemistry
Degree:
Doctor of Philosophy
Document Type:
Dissertation
Date of Defense:
September 29, 2005
Committee Members:
Christine Dolan Keating, Committee Chair/Co-Chair Andrew Ewing, Committee Member Erin Elizabeth Sheets, Committee Member William O Hancock, Committee Member
Keywords:
microcompartmentation aqueous two-phase system giant vesicle
Abstract:
Macromolecular crowding and microcompartmentation are fundamental features of the cell cytoplasm. Aqueous two-phase vesicles (ATPS GVs) are primitive “artificial cells” that exhibit both of these features and retain the activity of encapsulated proteins. I demonstrate dynamic organization of the aqueous compartment within ATPS GVs in response to small changes in temperature or osmotic pressure, the latter being useful for inducing asymmetric protein microcompartmentation. I demonstrate the use of colloidal gold scaffolds as a general method to dramatically increase protein partitioning within bulk aqueous two-phase systems (ATPS) with the retention of a significant level of activity. Additionally, I demonstrate complex temperature- and salt-dependent phase behavior of bulk ATPS and explain how this behavior can be rationalized through polymer hydration and conformation.
