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Understanding the role of charge on particle transport within semidilute and concentrated biopolymer solutions and tau protein condensates.

Date:
-
Location:
Zoom
Speaker(s) / Presenter(s):
Kanthi Nuti

Abstract: Biological polymer networks such as mucus, extracellular matrix, nuclear pore complex, and bacterial biofilms, play a critical role in governing the transport of nutrients, biomolecules and particles within cells and tissues. The interactions between particle and polymer chains are responsible for effective selective filtering of particles within these macromolecular networks. This selective filtering is not dictated by steric alone but must use additional interactions such electrostatics, hydrophobic and hydrodynamic effects to control particle transport within biogels. Depending on chemical composition and desired function, biogels use selective filtering to allow some particles to permeate while preventing others from penetrating the biogel. The mechanisms underlying selective filtering are still not well understood yet have important ramifications for a variety of biomedical applications. Controlling these non-steric interactions are critical to understanding molecular transport in vivo as well as for engineering optimized gel-penetrating therapeutics. Fluorescence correlation spectroscopy (FCS) is an ideal tool to study particle transport properties within uncharged and charged polymer solutions. In this dissertation, our research focuses primarily on the role of electrostatics on the particle diffusion behavior within polymer solutions in the semi-dilute and concentrated regimes.

Using a series of charged dye molecules, with similar size and core chemistry but varying net molecular charge, we systematically investigated their diffusion behavior in polymer solutions and networks made up of polysaccharide and proteins. Specifically, we studied in Chapter 3 the probe diffusion in semidilute and concentrated dextran solutions. The hindered diffusion observed in attractive gels is dependent on the probe net charge and shows a dependence on the solution ionic strength. Using a biotinylated probe, we also show evidence of an additional non-electrostatic interaction between the biotin molecule and the dextran polymer chains. In contrast, comparisons to a highly charged, water soluble polyvinylamine (PVAm) semidilute solution shows that all probes, regardless of charge, were highly hindered and a weaker dependence on solution ionic strength was observed. In Chapter 4, we characterized the transport properties of our probe molecules within pure and mixed charge solutions of amino(+)-dextran and carboxymethyl(-)-dextran. We show that these mixed charge polymer solutions still have the potential to be efficient filters for interacting particles even with comparably few attractive interaction sites. By chemical modification of the amino dextran, we also compare these results to those obtained in polyampholytic solutions. Lastly, we investigate the transport properties of both probes and a much larger bovine serum albumin (BSA) protein molecule within liquid-liquid phase separated (LLPS) tau protein in chapter 5. Tau is an intrinsically disordered protein with both positive and negatively charged amino acids. We show that despite having a high local protein concentration, tau droplets are relatively low density and comparable to semi-dilute polymer solutions. Both probe molecules and BSA are observed by FCS to be recruited within the liquid droplet resulting in ~10x fold increase in particle concentration inside the tau droplet compared to outside. Probe transport within the phase separated tau is sensitive to probe net charge and solution ionic strength. Lastly, we show that BSA transport inside the tau droplet can be fairly well described by using Stokes-Einstein adjusted for the experimentally determined microviscosity within the tau droplet.

 

Keywords: diffusion, biological gels, fluorescence correlation spectroscopy, electrostatic, interaction filtering.

 

Join the seminar at https://uky.zoom.us/j/9237836600

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