Intrinsically disordered proteins (IDPs) are a class of proteins lacking a fixed tertiary structure. IDPs are involved in a number of protein-misfolding diseases, such as cancers (α-Fetoprotein, p53), Alzheimer’s Disease (Aβ, tau protein), and Prion diseases, and study of their conformational changes is of significant biological relevance. Single molecule techniques can be utilized to bypass the inherent heterogeneity of IDPs in order to observe their individual conformations. Unfortunately, common techniques for studying biomolecules at the single-molecule level, namely confining the protein or tethering it to a surface, have been shown to affect the conformational dynamics of IDPs. The research presented shows the application of an Anti-Brownian Electrokinetic (ABEL) Trap which works to counteract a molecule’s Brownian motion and allow long-term study of biomolecules in solution without tethering them to a surface. We use time-resolved fluorescence anisotropy measurements which probe the global and local rotational dynamics of individual proteins – a measurement which would be nontrivially perturbed by tethering the IDPs to a surface – to ultimately ascertain information about the IDP’s conformations. Lastly, we use simulations of our data acquisition process to quantify the statistical robustness of our analysis.<br/>

Time-resolved anisotropy measurements of single, non-tethered tau proteins in solution

253rd National Meeting (2017)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.