The brain-specific tau protein binds directly through microtubules to regulate dynamically its structure and function. It also plays a critical role in the pathogenesis of a number of neurodegenerative disorders collectively known as tauopathies, the most common of which is Alzheimer’s disease (AD). Under pathological conditions, the natively unfolded tau protein self-assembles into filamentous structures of aggregated, hyperphosphorylated tau. In AD brains, tau accumulates in the neuronal perikarya and processes as paired helical filaments (PHF) forming the neurofibrillary tangles (NFT) characteristic of the disease. Prominent tau neurofibrillary pathology is a common feature in all tauopathies and its development is associated with progressive neuronal loss and cognitive decline. A precise understanding of the cellular, biochemical, and structural mechanisms involved in the process of tau protein aggregation and fibril formation is key to design strategies to prevent, slow down, or stop the neurodegenerative pathway leading to neuronal loss in AD and other tauopathies. Herein, we describe some complementary experimental procedures for PHF purification from human postmortem brain, tau expression and purification, as well as in vitro formation of tau filaments from purified recombinant tau.
Aggregation Alzheimer’s disease Neurofibrillary tangles Paired helical filaments Tau Tauopathies
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We would like to thank Dr. Jesús Avila for helpful discussions throughout this work. This project was supported by the Carlos III Institute of Health and the Spanish Ministry of Economy, Industry and Competitiveness (SAF2016-78603-R to MM).
Drechsel DN, Hyman AA, Cobb MH et al (1992) Modulation of the dynamic instability of tubulin assembly by the microtubule-associated protein tau. Mol Biol Cell 3(10):1141–1154CrossRefPubMedPubMedCentralGoogle Scholar
Grundke-Iqbal I, Iqbal K, Quinlan M et al (1986) Microtubule-associated protein tau. A component of Alzheimer paired helical filaments. J Biol Chem 261:6084–6089PubMedGoogle Scholar
Kosik KS, Joachim CL, Selkoe DJ (1986) Microtubule-associated protein tau (tau) is a major antigenic component of paired helical filaments in Alzheimer disease. Proc Natl Acad Sci U S A 83:4044–4048CrossRefPubMedPubMedCentralGoogle Scholar
Chen F, David D, Ferrari A et al (2004) Posttranslational modifications of tau – role in human tauopathies and modeling in transgenic animals. Curr Drug Targets 5(6):503–515CrossRefPubMedGoogle Scholar
Neve RL, Harris P, Kosik KS et al (1986) Identification of cDNA clones for the human microtubule-associated protein tau and chromosomal localization of the genes for tau and microtubule-associated protein 2. Brain Res 387:271–280PubMedGoogle Scholar
Lee G, Cowan N, Kirschner M (1988) The primary structure and heterogeneity of tau protein from mouse brain. Science 239:285–288CrossRefPubMedGoogle Scholar
Goedert M, Spillantini MG, Potier MC et al (1989) Cloning and sequencing of the cDNA encoding an isoform of microtubule-associated protein tau containing four tandem repeats: differential expression of tau protein mRNAs in human brain. EMBO J 8:393–399PubMedPubMedCentralGoogle Scholar
Hutton M, Lendon CL, Rizzu P et al (1998) Association of missense and 5′-splice-site mutations in tau with the inherited dementia FTDP-17. Nature 393:702–705CrossRefPubMedGoogle Scholar
Medina M, Hernández F, Avila J (2016) New features about tau function and dysfunction. Biomol Ther 6(2):E21Google Scholar
Medina M, Avila J (2014) New perspectives on the role of tau in Alzheimer’s disease. Implications for therapy. Biochem Pharmacol 88:540–547CrossRefPubMedGoogle Scholar
Goedert M, Jakes R (1990) Expression of separate isoforms of human tau protein: correlation with the tau pattern in brain and effects on tubulin polymerization. EMBO J 9:4425–4430Google Scholar
Greenberg SG, Davies P (1990) A preparation of Alzheimer paired helical filaments that displays distinct tau proteins by polyacrylamide gel electrophoresis. Proc Natl Acad Sci U S A 87(15):5827–5831CrossRefPubMedPubMedCentralGoogle Scholar
Sanbrook J, Frisch EF, Maniatis T (1989) Molecular cloning. A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
Gómez-Ramos A, Abad X, López Fanarraga M et al (2004) Expression of an altered form of tau in Sf9 insect cells results in the assembly of polymers resembling Alzheimer’s paired helical filaments. Brain Res 1007(1–2):57–64CrossRefPubMedGoogle Scholar
Pérez M, Valpuesta JM, Medina M et al (1996) Polymerization of tau into filaments in the presence of heparin: the minimal sequence required for tau-tau interaction. J Neurochem 67(3):1183–1190CrossRefPubMedPubMedCentralGoogle Scholar
Ksiezak-Reding H, Wall JS (2005) Characterization of paired helical filaments by scanning transmission electron microscopy. Microsc Res Tech 67(3–4):175–195Google Scholar