Details zur Publikation

Referenztyp Zeitschriften
DOI / URL Link
Titel (primär) The core of tau-paired helical filaments studied by scanning transmission electron microscopy and limited proteolysis
Autor von Bergen, M.; Barghorn, S.; Müller, S.A.; Pickhardt, M.; Biernat, J.; Mandelkow, E.-M.; Davies, P.; Aebi, U.; Mandelkow, E.;
Journal / Serie Biochemistry
Erscheinungsjahr 2006
Department PROTEOM;
Band/Volume 45
Heft 20
Sprache englisch;
Abstract In Alzheimer's disease and frontotemporal dementias the microtubule-associated protein tau forms intracellular paired helical filaments (PHFs). The filaments formed in vivo consist mainly of full-length molecules of the six different isoforms present in adult brain. The substructure of the PHF core is still elusive. Here we applied scanning transmission electron microscopy (STEM) and limited proteolysis to probe the mass distribution of PHFs and their surface exposure. Tau filaments assembled from the three repeat domain have a mass per length (MPL) of ~60 kDa/nm and filaments from full-length tau (htau40K280 mutant) have ~160 kDa/nm, compared with ~130 kDa/nm for PHFs from Alzheimer's brain. Polyanionic cofactors such as heparin accelerate assembly but are not incorporated into PHFs. Limited proteolysis combined with N-terminal sequencing and mass spectrometry of fragments reveals a protease-sensitive N-terminal half and semiresistant PHF core starting in the first repeat and reaching to the C-terminus of tau. Continued proteolysis leads to a fragment starting at the end of the first repeat and ending in the fourth repeat. PHFs from tau isoforms with four repeats revealed an additional cleavage site within the middle of the second repeat. Probing the PHFs with antibodies detecting epitopes either over longer stretches in the C-terminal half of tau or in the fourth repeat revealed that they grow in a polar manner. These data describe the physical parameters of the PHFs and enabled us to build a model of the molecular arrangement within the filamentous structures.
ID 3093
dauerhafte UFZ-Verlinkung https://www.ufz.de/index.php?en=20939&ufzPublicationIdentifier=3093
von Bergen, M., Barghorn, S., Müller, S.A., Pickhardt, M., Biernat, J., Mandelkow, E.-M., Davies, P., Aebi, U., Mandelkow, E. (2006):
The core of tau-paired helical filaments studied by scanning transmission electron microscopy and limited proteolysis
Biochemistry 45 (20), 6446 - 6457