title: Oligomer formation of tau protein hyperphosphorylated in cells.
abstract: Abnormal phosphorylation ("hyperphosphorylation") and aggregation of Tau protein are hallmarks of Alzheimer disease and other tauopathies, but their causative connection is still a matter of debate. Tau with Alzheimer-like phosphorylation is also present in hibernating animals, mitosis, or during embryonic development, without leading to pathophysiology or neurodegeneration. Thus, the role of phosphorylation and the distinction between physiological and pathological phosphorylation needs to be further refined. So far, the systematic investigation of highly phosphorylated Tau was difficult because a reliable method of preparing reproducible quantities was not available. Here, we generated full-length Tau (2N4R) in Sf9 cells in a well defined phosphorylation state containing up to ∼20 phosphates as judged by mass spectrometry and Western blotting with phospho-specific antibodies. Despite the high concentration in living Sf9 cells (estimated ∼230 μm) and high phosphorylation, the protein was not aggregated. However, after purification, the highly phosphorylated protein readily formed oligomers, whereas fibrils were observed only rarely. Exposure of mature primary neuronal cultures to oligomeric phospho-Tau caused reduction of spine density on dendrites but did not change the overall cell viability.
authors.affiliation: From the DZNE, German Center for Neurodegenerative Diseases, 53175 Bonn, Germany, the CAESAR Research Center, 53175 Bonn, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Tepper
authors.affiliation: From the DZNE, German Center for Neurodegenerative Diseases, 53175 Bonn, Germany, the CAESAR Research Center, 53175 Bonn, Germany.
authors.forename: Jacek
authors.initials: J
authors.lastname: Biernat
authors.affiliation: From the DZNE, German Center for Neurodegenerative Diseases, 53175 Bonn, Germany, the CAESAR Research Center, 53175 Bonn, Germany.
authors.forename: Satish
authors.initials: S
authors.lastname: Kumar
authors.affiliation: the Department of Biosystems Science and Engineering, ETHZ, 4058 Basel, Switzerland.
authors.forename: Susanne
authors.initials: S
authors.lastname: Wegmann
authors.affiliation: the Institute of Biochemistry, Faculty of Medicine, Justus-Liebig-University, 35390 Giessen, Germany, and.
authors.forename: Thomas
authors.initials: T
authors.lastname: Timm
authors.affiliation: From the DZNE, German Center for Neurodegenerative Diseases, 53175 Bonn, Germany, the CAESAR Research Center, 53175 Bonn, Germany.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Hübschmann
authors.affiliation: the Joint Laboratory for Structural Biology of Infection and Inflammation, University of Hamburg and University of Lübeck, ℅DESY, 22603 Hamburg, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Redecke
authors.affiliation: From the DZNE, German Center for Neurodegenerative Diseases, 53175 Bonn, Germany, the CAESAR Research Center, 53175 Bonn, Germany.
authors.forename: Eva-Maria
authors.initials: EM
authors.lastname: Mandelkow
authors.affiliation: the Department of Biosystems Science and Engineering, ETHZ, 4058 Basel, Switzerland.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Müller
authors.affiliation: From the DZNE, German Center for Neurodegenerative Diseases, 53175 Bonn, Germany, the CAESAR Research Center, 53175 Bonn, Germany, Eckhard.Mandelkow@dzne.de.
authors.forename: Eckhard
authors.initials: E
authors.lastname: Mandelkow
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D045163:Antibodies, Phospho-Specific / Q000737:chemistry; D016367:Baculoviridae / Q000235:genetics; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D015870:Gene Expression; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010449:Peptide Mapping; D010766:Phosphorylation; D061251:Primary Cell Culture; D066329:Protein Aggregates; D055503:Protein Multimerization / Q000235:genetics*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D061987:Sf9 Cells; D018411:Spodoptera; D016875:tau Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology
chemical_list: D045163:Antibodies, Phospho-Specific; D066329:Protein Aggregates; D011994:Recombinant Proteins; D016875:tau Proteins
keywords: Fluorescence Anisotropy; Neuron; Oligomers; Phosphorylation; Synapse; Tau Protein (Tau); Time-correlated Single Photon Counting; Toxicity