Abstract
A native conformation of a protein is essential for its biological role. In certain conditions, some proteins show non-native conformations, leading to aggregation, which in turn may produce severe pathologies. Such physiological conditions are classified as protein misfolding diseases. Alzheimer’s disease (AD) is the most common form of dementia. Extracellular senile plaques formed by Amyloid β and intracellular aggregates formed by microtubule-associated protein Tau (MAPT) are the hallmarks of AD. Physiological role of MAPT is to maintain the integrity and stability of microtubules, however it tends to self-aggregate forming intracellular paired helical filaments (PHFs) during AD. MAPT is also subjected to various post-translational modifications such as phosphorylation, glycosylation, truncation, and acetylation. Being natively unfolded, MAPT is prone to full characterization at atomic level. Small-angle X-ray scattering (SAXS) is often applied in combination with other biophysical methods, like nuclear magnetic resonance (NMR), circular dichroism (CD), fluorescence spectroscopy, analytical ultracentrifugation (AUC), or dynamic light scattering (DLS) to characterize natively unfolded systems. Here we describe the practical aspects of MAPT characterization by SAXS and CD in detail as well as outline the inferred structural and functional implications.
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Acknowledgments
The authors thank Dr. Hirekodathakallu V. Thulasiram (CSC0130) for his excellent Molecular Biology Laboratory (MBL) facility at the CSIR-NCL, Pune. Tau constructs were kindly gifted by Prof. Roland Brandt from University of Osnabruck, Germany and Prof. Jeff Kuret from Ohio State University College of Medicine, USA. N.V.G. acknowledges research fellowship from University of Grant Commission (UGC) by government of India. This project is supported in part by grants from the DST-SERB SB/YS/LS-355/2013 and 12th 5-year plan CSIR-Network Project BSC0115. The authors acknowledge Dr. Ranjan Singh, Shweta Sonawane, Dr. Yann Stercks, and Dr. Ritika Sethi for proofreading the manuscript and for useful comments. The authors have declared no conflict of interest.
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Gorantla, N.V., Shkumatov, A.V., Chinnathambi, S. (2017). Conformational Dynamics of Intracellular Tau Protein Revealed by CD and SAXS. In: Smet-Nocca, C. (eds) Tau Protein. Methods in Molecular Biology, vol 1523. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6598-4_1
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