Page 13 - T. Protein modification and regulation

Core sequence of Tau regulating its aggregation process

Jeeyoung Lee, Ly Thi Huong Luu Le, Min Jae Lee
Biochemisty and Molecular Biology, Seoul National University College of Medicine, Seoul,
Korea

BACKGROUND AIM

In tauopathies, the microtubule-associated protein tau (Tau) is To examine whether the protofilament core sequence
abnormally aggregated and forms neurofibrillary tangles (NFTs). affects the aggregation process of Tau FL, we purified the
A recent study using human AD brains revealed the core recombinant Tau core protein and raised its targeting
sequence (Tau core) of the ordered b-sheet structure. Among
the four microtubule-binding repeat domains (MTBRs) in the full- antibodies. Our in vitro oligomerization assay, electron
length Tau (Tau FL), the Tau core consists of V306 to F378 of microscopic analysis, and dot blot assay indicated that Tau
Core facilitates
Tau FL, spanning MTBRs 3 and 4.
METHODS

His-tagged human tau was then purified on Ä KTA pure using a HiTrap TALON crude column. Proteins were eluted with 150 mM
imidazole (50 mM NaH2PO4 [pH 7.4], 300 mM NaCl, and 150 mM imidazole). The eluted tau protein was supplemented with
dithiothreitol (DTT) to a final concentration of 0.1 mM for reducing disulfide bonds and was stored at −80 °C. Tau core proteins
were purified by the same protocol as Tau FL. Tau FL proteins were incubated with Tau core in the absence of any aggregation
inducers.
RESULTS

CONCLUSION REFERENCES
• Fitzpatrick AWP et al, Nature. 2017 Jul 13; 547(7662):185-190.
The Tau aggregates induced by the Tau core form a
thicker and shorter filament than the Tau aggregates • Chen HH et al, Sci Rep. 2018 Nov 13;8(1): 16725.
by the heparin. This form of filament is a form known to • Zhang W et al, Elife. 2019 Feb 5;8. pii: e43584.
be more toxic by recent studies. Therefore, the Tau
filament due to the Tau core derived from the Tau FL Contact information
sequence is expected to be closer to the type of
filament that can be found in vivo.
JY Lee : onnu2@snu.ac.kr / MJ Lee : minjlee@snu.ac.kr