STOCHASTIC BIOPHYSICAL INTERACTIONS WITHIN AQUAPORIN-4 ASSEMBLIES
Abstract
Fundamental biological principles are often based on direct observation and this also holds true for much of physics and chemistry. The power of direct observation lays in the fact that is proof positive that something exists and can act in a certain manner. Many observations lead to generalizations and hypotheses that can be tested in much greater detail. Only recently has it been possible to directly observe single molecules and proteins as they move, interact, and react with their surroundings. In this one-year collaborative feasibility study we will use recently developed biochemical and bioengineering tools developed at the University of Bari (Italy) along with recently developed single protein tracking instrumentation and bioanalysis at Washington State University to make a movie – actually many movies – of AQP4 assembly and disassemble as a function of AQP4 isoform, environment and temperature. The individual observations will be cast into a generalized framework model that includes side trajectories and dead ends – these often carry a significant amount of entropic weight and have a regulatory function. This framework model will serve as a working hypothesis of AQP4 self-assembly and average measurements will help determine most probable paths within the framework and free energy driving forces (through a kinetic analysis). Temperature changes will be used to estimate enthalpy and entropy changes between states within the framework. Project goals will be met though four major objectives; (1) establishment of a recent international collaboration and interaction between the UNIBA (Italy) and Washington State University, (2) build a single protein tracking experiment to follow individual AQP4s over long periods of time, (3) observe AQP4 assemblies in expression systems and in brain astrocytes by super-resolution microscopy, and (4) understand the stochastic biophysical properties of AQP4 self-assembly / disassembly through direct observation.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Aug 12, 2021
- Source ID
- FA95502010324
Entities
People
- James A Brozik
Organizations
- Air Force Office of Scientific Research
- United States Air Force
- Washington State University