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@PHDTHESIS{Chinnathambi:270141,
author = {Chinnathambi, Subash},
title = {{I}nfluence of {P}hosphorylation on the {P}athological
{C}onformation of {H}uman {M}icrotubule {A}ssociated
{P}rotein {T}au},
school = {Universität Hamburg},
type = {Dissertation},
reportid = {DZNE-2024-00741},
pages = {159 pages},
year = {2011},
note = {Dissertation, Universität Hamburg, 2011},
abstract = {Tau is one of the microtubule associated proteins which can
bind and stabilize microtubules. It is classified as a
natively unfolded protein. It has a very low content of
secondary structural elements. Tau aggregates into insoluble
fibers called paired helical filaments in Alzheimer’s
disease. The residual folding and intramolecular
interactions within tau are largely unknown, and the factors
contributing to the formation and stability of PHFs and
different morphologies of PHFs are not well understood.The
results of the present studies are aimed at better
understanding of the effect of phosphorylation on the
aggregation propensity and tau microtubule interaction.The
effect of phosphorylation sites in the KXGS motifs within
the repeats and in the SP/TP motifs of the flanking regions
were studied. We also studied tau fibrilpolymorphism by
Electron Microscopy and Atomic Force Microscopy. We further
studied tau in solution by increasing temperature using
different biophysical methods (CD spectroscopy, static and
dynamic light scattering and small angle x-ray scattering).
These observations could form a basis for elucidating the
molecular and physiological role of phosphorylation at the
repeat domains and in the flanking regions. The results are
summarized as follows(i) The pseudo-phosphorylation of KXGE
motifs within the repeat domains of tau has a strong
negative effect on both the tau-microtubule interaction and
the aggregation into paired helical filaments. KXGE mutants
on the basis of three repeat domain and four repeat domain
constructs show a low tendency to aggregate and form
amorphous aggregates instead of PHFs. Pseudo-phosphorylation
in the SP/TP motifs in the N- and C-terminal domains of the
flanking regions has only a moderate effect, both on the
tau-microtubule interaction and on the aggregation into
paired helical filaments. Overall, the results show that the
physiological tau-microtubule interaction and the
pathological tau-tau interaction similarly can be influenced
by phosphorylation.(ii) The combined pseudo-phosphorylation
of AT8* and PHF1 sites leads to compaction of the paperclip
conformation, such that the N-terminus approaches the repeat
domain. When the Cterminus tail is cleaved from tau, the
compaction becomes even stronger by combining
pseudophosphorylated AT8*, AT100, and PHF1 epitopes. This is
accompanied by a strong increase in the reaction with
conformation-dependent antibodies MC1 and Alz-50, suggesting
the generation of a pathological conformation characteristic
of tau in AD. Furthermore, the compact paperclip
conformation enhances the aggregation propensity to paired
helical filaments but has little effect on microtubule
interactions. Discussion 89(iii) High resolution atomic
force and electron microscopy were employed to study
fibrilsassembled from different human tau isoforms and
domains. All fibrils reveal structuralpolymorphism: the
“thin twisted” and “thin smooth” fibrils resemble
flat ribbons(cross-section ~10 x 15 nm) with diverse twist
periodicities. “Thick fibrils” show periodicities of
~65-70 nm and thicknesses of ~9-18 nm as routinely reported
for“paired helical filaments” but structurally resemble
heavily twisted ribbons.Therefore, thin and thick fibrils
assembled from different human tau isoforms challenge
current structural models of paired helical filaments. The
full-length tau and repeat domain constructs assemble into
fibrils of similar thickness, the “fuzzy coat” of tau
protein termini surrounding the fibril axis is nearly
invisible for atomicforce microscopy and electron
microscopy, presumably because of its high flexibility.(iv)
The unfolded nature of tau protein allows it to adopt either
more extended or compact conformations. It is intriguing to
speculate that the compaction and memory effect observed
after rapid heating/cooling may be related to the compaction
observed upon hyperphosphorylation, characteristic of
incipient neuronal degeneration in AD. The “memory
effect” is observed with full-length tau, but not with the
repeat domain alone, suggesting that the interplay between
domains in the whole protein might be responsible for the
effect. One possible explanation could be the interplay
between the acidic N-terminal domain (which varies among the
tau isoforms due to alternative splicing) and the basic
repeat domain (which also differs between the isoforms).},
cin = {AG Mandelkow 1},
cid = {I:(DE-2719)1013014},
pnm = {899 - ohne Topic (POF4-899)},
pid = {G:(DE-HGF)POF4-899},
typ = {PUB:(DE-HGF)11},
urn = {urn:nbn:de:gbv:18-56123},
url = {https://pub.dzne.de/record/270141},
}