% IMPORTANT: The following is UTF-8 encoded. This means that in the presence
% of non-ASCII characters, it will not work with BibTeX 0.99 or older.
% Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or
% “biber”.
@INBOOK{Hochmair:165615,
author = {Hochmair, Janine and Exner, Christian and Betzel, Christian
and Mandelkow, Eckhard and Wegmann, Susanne},
title = {{L}ight {M}icroscopy and {D}ynamic {L}ight {S}cattering to
{S}tudy {L}iquid-{L}iquid {P}hase {S}eparation of {T}au
{P}roteins {I}n {V}itro.},
volume = {2551},
address = {New York, NY},
publisher = {Springer US},
reportid = {DZNE-2022-01748},
isbn = {978-1-0716-2596-5 (print)},
series = {Methods in Molecular Biology},
pages = {225 - 243},
year = {2023},
comment = {Protein Aggregation / Cieplak, Andrzej Stanisław (Editor)
; New York, NY : Springer US, 2023, Chapter 15 ; ISSN:
1064-3745=1940-6029 ; ISBN:
978-1-0716-2596-5=978-1-0716-2597-2 ;
doi:10.1007/978-1-0716-2597-2},
booktitle = {Protein Aggregation / Cieplak, Andrzej
Stanisław (Editor) ; New York, NY :
Springer US, 2023, Chapter 15 ; ISSN:
1064-3745=1940-6029 ; ISBN:
978-1-0716-2596-5=978-1-0716-2597-2 ;
doi:10.1007/978-1-0716-2597-2},
abstract = {Tau is an intrinsically disordered protein that binds and
stabilizes axonal microtubules (MTs) in neurons of the
central nervous system. The binding of Tau to MTs is
mediated by its repeat domain and flanking proline-rich
domains. The positively charged (basic) C-terminal half of
Tau also mediates the assembly Tau into fibrillar aggregates
in Alzheimer's disease (AD) and tauopathy brains. In recent
years, another assembly form of Tau has been identified: Tau
can undergo liquid-liquid phase separation (LLPS), which
leads to its condensation into liquid-dense phases, either
by complex coacervation with polyanions like heparin or RNA
or through 'self-coacervation' at high Tau concentrations.
Condensation of Tau in the absence of polyanions can be
enhanced by the presence of molecular crowding agents in a
dilute Tau solution. In vitro experiments using recombinant
purified Tau are helpful to study the physicochemical
determinants of Tau LLPS, which can then be extrapolated
into the cellular context. Tau condensation is a new aspect
of Tau biology that may play a role for the initiation of
Tau aggregation, but also for its physiological function(s),
for example, the binding to microtubules. Here we describe
how to study the condensation of Tau in vitro using light
microscopy, including fluorescence recovery after
photobleaching (FRAP), to assess the shape and molecular
diffusion in the condensates, a proxy for the degree of
condensate percolation. We also describe turbidity
measurements of condensate-containing solutions to assess
the overall amount of LLPS and time-resolved dynamic light
scattering (trDLS) to study the formation and size of Tau
condensates.},
keywords = {Humans / tau Proteins: metabolism / Microscopy / Dynamic
Light Scattering / Alzheimer Disease: metabolism /
Coacervation (Other) / Condensation (Other) / Crowding
agents (Other) / FRAP (Other) / LLPS (Other) / MAPT (Other)
/ Polyethylene glycol (Other) / RNA (Other) / tau Proteins
(NLM Chemicals) / polyanions (NLM Chemicals)},
cin = {AG Wegmann / AG Mandelkow 1},
ddc = {570},
cid = {I:(DE-2719)1810006 / I:(DE-2719)1013014},
pnm = {352 - Disease Mechanisms (POF4-352)},
pid = {G:(DE-HGF)POF4-352},
typ = {PUB:(DE-HGF)7},
pubmed = {pmid:36310206},
doi = {10.1007/978-1-0716-2597-2_15},
url = {https://pub.dzne.de/record/165615},
}