Characterization of intrinsically disordered proteins
Abstract
Intrinsically disordered proteins (IDPs) or proteins containing disordered regions (IDRs) are interesting both biophysically and physiologically, but remain difficult to study by current methods in structural biology and biophysics. IDPs and IDRs cannot be described by a single representative structure, but are best characterized by ensembles of conformations that are distinctly different from a random coil. The regulatory domain (RD) of human tyrosine hydroxylase (TyrH) is an example of hybrid protein containing structured and IDR region. TyrH is regulated by phosphorylation of RD-TyrH within the IDR and subsequent interaction with 14-3-3 proteins. As an example of IDPs for this proposal we selected the microtubule associated protein 2c (Map2c) and its homologous tau protein containing highly similar C-terminal disordered regions. Yet, the experimentally observed phosphorylation pattern is very different for the two IDPs, suggesting that residual structural preferences govern the phosphorylation propensity. We will characterize the conformational ensembles of RD-TyrH, Map2c and tau. We hypothesize that shifts in the conformational ensembles of these IDPs/IDRs are crucial to understand their function and will further develop integrative experimental and computational approaches to characterize these shifts. We will study our model proteins using a wide range of techniques to characterize the shifts in the conformational ensembles. Computer simulations and nuclear magnetic resonance (NMR) spectroscopy are key techniques to study IDPs/IDRs at an atomic resolution. The applicability of both methods is limited by the potential size of the conformational ensembles. We will develop and merge the necessary methodologies. Computationally, we will use enhanced sampling of backbone degrees of freedom of fragments and merge these fragments to construct a large number of viable IDP conformations. Experimental data from NMR will be used to guide the formation of representative ensembles. Innovation The direct interplay of experimental and computational techniques is the key point of this proposal. While both approaches individually cannot be expected to understand the function of IDPs at a detailed level, their combination will. We will develop novel enhanced sampling methods to generate realistic conformations and use the experimental data to form realistic ensembles. We bring together two groups with highly complementing expertises in both areas.
Publikationen
Project staff
Chris Oostenbrink
Univ.Prof. Dr. Chris Oostenbrink
chris.oostenbrink@boku.ac.at
Tel: +43 1 47654-89401, 89411, 89419
Project Leader
01.05.2020 - 30.04.2023