bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2023,
Номер
unknown
Опубликована: Дек. 2, 2023
ABSTRACT
Immunofluorescence
is
a
common
method
to
localise
proteins
within
their
cellular
context
via
fluorophore
labelled
antibodies
and
for
some
applications
without
alternative.
However,
protein
targets
evade
detection
due
low
abundance
or
accessibility
issues.
In
addition,
imaging
methods
require
massive
reduction
in
antigen
density
thus
impeding
of
even
medium-abundant
proteins.
Here,
we
show
that
the
fusion
target
TurboID,
biotin
ligase
labelling
lysine
residues
close
proximity,
subsequent
biotinylation
by
fluorescent
streptavidin
offers
an
“all
one”
solution
above-mentioned
restrictions.
For
wide
range
tested,
signal
was
significantly
stronger
than
antibody
signal,
markedly
improving
sensitivity
expansion
microscopy
correlative
light
electron
microscopy,
with
no
loss
resolution.
Importantly,
phase-separated
regions,
such
as
central
channel
nuclear
pores,
nucleolus
RNA
granules,
were
readily
detected
streptavidin,
while
most
fail
label
these
environments.
When
TurboID
used
tandem
HA
epitope
tag,
co-probing
anti-HA
can
be
map
antibody-
certain
regions.
As
proof
principle,
mapped
access
all
trypanosome
pore
(NUPs)
found
restricted
FG
NUPs
are
known
phase-separated,
non-FG
Nups
could
labelled.
Lastly,
resolve
dynamic,
temporally
spatially
distinct
sub-complexes
and,
specific
cases,
reveal
history
dynamic
interaction.
conclusion,
has
major
advantages
lowly
abundant
inaccessible
provide
information
on
interactions
biophysical
environment.
Journal of Cellular Biochemistry,
Год журнала:
2024,
Номер
125(3)
Опубликована: Фев. 13, 2024
Abstract
When
the
SARS‐CoV‐2
virus
infects
humans,
it
leads
to
a
condition
called
COVID‐19
that
has
wide
spectrum
of
clinical
manifestations,
from
no
symptoms
acute
respiratory
distress
syndrome.
The
initiates
damage
by
attaching
ACE‐2
protein
on
surface
endothelial
cells
line
blood
vessels
and
using
these
as
hosts
for
replication.
Reactive
oxygen
species
levels
are
increased
during
viral
replication,
which
oxidative
stress.
About
three‐fifths
(~60%)
people
who
get
infected
with
eradicate
their
body
after
28
days
recover
normal
activity.
However,
large
fraction
(~40%)
suffer
various
(anosmia
and/or
ageusia,
fatigue,
cough,
myalgia,
cognitive
impairment,
insomnia,
dyspnea,
tachycardia)
beyond
12
weeks
diagnosed
syndrome
long
COVID.
Long‐term
studies
in
group
contracted
have
been
contrasted
noninfected
matched
people.
A
subset
can
be
distinguished
set
cytokine
markers
persistent,
low‐grade
inflammation
often
self‐report
two
or
more
bothersome
symptoms.
No
medication
alleviate
efficiently.
Coronavirus
nucleocapsid
proteins
investigated
extensively
potential
drug
targets
due
key
roles
among
is
ability
bind
respective
genomic
RNAs
incorporation
into
emerging
virions.
This
review
highlights
basic
its
undergo
liquid–liquid
phase
separation.
We
hypothesize
this
separation
may
contribute
hypothesis
unlocks
new
investigation
angles
could
potentially
open
novel
avenues
better
understanding
COVID
treating
condition.
Nature Communications,
Год журнала:
2024,
Номер
15(1)
Опубликована: Окт. 9, 2024
Parkinson's
disease
(PD)
and
Dementia
with
Lewy
Bodies
(DLB)
are
neurodegenerative
disorders
characterized
by
the
accumulation
of
α-synuclein
aggregates.
forms
droplets
via
liquid-liquid
phase
separation
(LLPS),
followed
liquid-solid
(LSPS)
to
form
amyloids,
how
this
process
is
physiologically-regulated
remains
unclear.
β-synuclein
colocalizes
in
presynaptic
terminals.
Here,
we
report
that
partitions
into
condensates
promotes
LLPS,
slows
down
LSPS
α-synuclein,
while
disease-associated
mutations
lose
these
capacities.
Exogenous
improves
movement
defects
prolongs
lifespan
an
α-synuclein-expressing
NL5901
Caenorhabditis
elegans
strain,
mutants
aggravate
symptoms.
Decapeptides
targeted
at
α-/β-synuclein
interaction
sites
rationally
designed,
which
suppress
rescue
defects,
prolong
C.
NL5901.
Together,
unveil
a
Yin-Yang
balance
between
α-
underlying
normal
states
PD
DLB
therapeutical
potentials.
Immunofluorescence
localises
proteins
via
fluorophore-labelled
antibodies.
However,
some
evade
detection
due
to
antibody-accessibility
issues
or
because
they
are
naturally
low
abundant
antigen
density
is
reduced
by
the
imaging
method.
Here,
we
show
that
fusion
of
target
protein
biotin
ligase
TurboID
and
subsequent
biotinylation
fluorescent
streptavidin
offers
an
‘all
in
one’
solution
these
restrictions.
For
all
tested,
signal
was
significantly
stronger
than
antibody
signal,
markedly
improving
sensitivity
expansion
microscopy
correlative
light
electron
microscopy.
Importantly,
within
phase-separated
regions,
such
as
central
channel
nuclear
pores,
nucleolus,
RNA
granules,
were
readily
detected
with
streptavidin,
while
most
antibodies
failed.
When
used
tandem
HA
epitope
tag,
co-probing
anti-HA
can
map
created
a
for
trypanosome
pore.
Lastly,
resolves
dynamic,
temporally,
spatially
distinct
sub-complexes
and,
specific
cases,
reveals
history
dynamic
interaction.
In
conclusion,
has
major
advantages
lowly
inaccessible
addition,
provides
information
on
interactions
biophysical
environment.
Signal Transduction and Targeted Therapy,
Год журнала:
2024,
Номер
9(1)
Опубликована: Ноя. 18, 2024
Abstract
Once
considered
unconventional
cellular
structures,
membraneless
organelles
(MLOs),
substructures
involved
in
biological
processes
or
pathways
under
physiological
conditions,
have
emerged
as
central
players
dynamics
and
function.
MLOs
can
be
formed
through
liquid-liquid
phase
separation
(LLPS),
resulting
the
creation
of
condensates.
From
neurodegenerative
disorders,
cardiovascular
diseases,
aging,
metabolism
to
cancer,
influence
on
human
health
disease
extends
widely.
This
review
discusses
underlying
mechanisms
LLPS,
biophysical
properties
that
drive
MLO
formation,
their
implications
for
We
highlight
recent
advances
understanding
how
physicochemical
environment,
molecular
interactions,
post-translational
modifications
regulate
LLPS
dynamics.
offers
an
overview
discovery
current
biomolecular
condensate
conditions
diseases.
article
aims
deliver
latest
insights
by
analyzing
research,
highlighting
critical
role
organization.
The
discussion
also
covers
membrane-associated
condensates
cell
signaling,
including
those
involving
T-cell
receptors,
stress
granules
linked
lysosomes,
within
Golgi
apparatus.
Additionally,
potential
targeting
clinical
settings
is
explored,
promising
avenues
future
research
therapeutic
interventions.
Journal of Chemical Information and Modeling,
Год журнала:
2025,
Номер
unknown
Опубликована: Март 7, 2025
Chronic
traumatic
encephalopathy
(CTE)
is
a
unique
tauopathy
mostly
diagnosed
in
contact
sports
athletes,
such
as
those
active
American
football,
boxing,
soccer,
etc.
The
hyperphosphorylated
fibrillar
aggregates
composed
of
self-assembled
tau
protein
are
the
pathological
hallmark
CTE,
and
inhibiting
aggregation
or
disassociating
has
been
considered
promising
avenue
to
prevent
treat
CTE.
Caffeine
(CA)
well-known
psychostimulant
can
be
found
coffee,
tea,
soft
drinks.
In
vitro
experiments
revealed
that
CA
could
effectively
inhibit
wild-type
disassemble
preformed
fibrils.
However,
atomic
effect
underlying
molecular
mechanisms
remain
largely
elusive.
this
study,
we
performed
multitude
replica
exchange
with
solute
tempering
2
(REST2)
conventional
dynamics
(CMD)
simulations
43.8
μs
total
on
models
without
CA,
including
third
fourth
microtubule-binding
repeats
(R3-R4)
monomer
CTE-related
R3-R4
protofibril
fibril.
results
prominently
β-sheet
formation
disrupt
structure
protofibril,
inducing
adopt
loosely
packed
extended
conformations.
H-bonding
π-π
stacking
interactions
drove
binding
monomer,
while
hydrophobic
made
an
extra
contribution
protofibril.
Strikingly,
stably
bind
cavity
which
might
occupy
space
entering
cofactor.
What
more,
destabilized
fibril
played
role
reversing
liquid-to-solid
phase
transition
(LSPT)
tau.
Our
study
systematically
uncovered
atomic-level
aggregation,
offers
theoretical
foundation
for
design
drugs
