LEA_4 motifs function alone and in conjunction with synergistic cosolutes to protect a labile enzyme during desiccation
Protein Science,
Год журнала:
2025,
Номер
34(2)
Опубликована: Янв. 22, 2025
Abstract
Organisms
from
all
kingdoms
of
life
depend
on
Late
Embryogenesis
Abundant
(LEA)
proteins
to
survive
desiccation.
LEA
are
divided
into
broad
families
distinguished
by
the
presence
family‐specific
motif
sequences.
The
LEA_4
family,
characterized
11‐residue
motifs,
plays
a
crucial
role
in
desiccation
tolerance
numerous
species.
However,
these
motifs
function
is
unclear,
with
some
studies
finding
that
they
recapitulate
full‐length
vivo,
and
other
opposite
result.
In
this
study,
we
characterize
ability
protect
desiccation‐sensitive
enzyme,
citrate
synthase
(CS),
loss
during
We
show
here
not
only
prevent
CS
but
also
can
do
so
more
robustly
via
synergistically
interactions
cosolutes.
Our
analysis
further
suggests
cosolutes
induce
synergy
manner
correlates
transfer
free
energy.
This
research
advances
our
understanding
demonstrating
their
specific
clients
varying
degrees
protective
capacity
modulated
chemical
environment.
findings
extend
beyond
realm
tolerance,
offering
insights
interplay
between
IDPs
By
investigating
highlight
broader
strategies
for
protein
stability
function.
Язык: Английский
Diversity in the protective role(s) of the conserved motif 1 from tardigrade cytoplasmic‐abundant heat‐soluble proteins during drying
Protein Science,
Год журнала:
2025,
Номер
34(3)
Опубликована: Фев. 19, 2025
In
their
recent
manuscript,
Protective
roles
of
highly
conserved
motif
1
in
tardigrade
cytosolic-abundant
heat
soluble
protein
extreme
environments,
Kang
et
al.
(2024)
investigate
the
putative
role
a
cytoplasmic-abundant
heat-soluble
(CAHS)
protein,
PrCAHS
1,
desiccation
protection.
To
do
so,
test
ability
different
regions
to
confer
protection
desiccation-sensitive
enzyme
lactate
dehydrogenase
(LDH).
doing
this,
they
compare
how
mixtures
with
mass
ratios
LDH
and
retain
activity
after
drying
rehydration.
However,
another
way
comparing
protective
capacity
protectants
molecular
weights
is
at
similar
molar
ratios.
Since
19-mer
has
weight
that
approximately
11
times
lower
than
full-length
choices
about
comparisons
(weight
vs.
comparisons)
are
made
could
influence
outcome
comparison.
This
led
us
reanalyze
data
for
converting
reported
ratios,
performing
statistical
analysis
across
(Figure
1a,b).
reanalysis
confirmed
finding
by
outperforms
protecting
during
drying.
this
result
was
surprising
us,
since
other
proteins
from
CAHS
family
provide
robust
(Biswas
al.,
2024;
Boothby
2017;
Hesgrove
2021;
Kc
Nguyen
2022;
Packebush
2023;
Piszkiewicz
2019;
Sanchez-Martinez
2024)
so
more
efficiently
individual
domains
2023).
observation
raises
question
whether
feature
among
proteins.
assess
if
1s
proteins,
we
performed
set
assay
those
(2024),
using
an
ortholog
HeCAHS
8,
derived
8.
Here
use
8
behave
similarly
1.
We
observed
opposite
obtained
where
provided
LDH,
whereas
did
not
1c,d).
From
these
results,
one
can
infer
while
some
may
help
enzymatic
greater
it
from,
sequences
not.
suggests
functional
diversity
rather
all
These
results
line
study
found
LEA_4
robustly
protect
drying,
motifs
(Kc
Nicholson
2025).
Why
might
have
were
al
(2024)?
One
possibility
distinct
general,
truly
function
differently.
possibility,
fact
sequence
1e),
minor
differences
composition
account
differential
between
two
motifs.
Previous
work
demonstrated
small
changes
CAHS-inspired
peptides
lead
large
structural
ensemble
Giubertoni
2024),
which
turn
affect
mediators
2024).
Another
possible
source
discrepancy
methods
measuring
activity.
While
indirect
tetrazolium/formazan
assay,
opted
method
directly
assays
presence
NADH
(a
primary
product
activity),
widely
used
tolerance
field
Goyal
2005;
Although
methodology
as
been
utilized
former
on
LEA
protein's
(Hatanaka
2013),
does
necessarily
make
best
approach
assay.
Several
issues
exist
assays,
including
propensity
formazan
precipitate
high
concentration,
thus
causing
major
inconsistencies
quantification
(Riss
2016).
The
effect
such
precipitation
concentrations
sample
levels
appear
actually
low
Companies
sell
often
mention
include
excipients
reduce
precipitation;
however,
generally
difficult
obtain
precise
information
what
effectiveness
preventing
precipitation.
explanation
why
be
suggested
or
domains,
prevent
aggregation
LDH.
anti-aggregation
potential
measure
absorbance
340
nm
solutions
prepared
desiccating
then
rehydrating
alone
authors
first
+
1f,
replotted
They
conclude
latter
solution
alone,
increased
due
but
itself,
contribution
readings
impossible
based
data.
It
worth
mentioning
here
known
undergo
phase
transition
form
gels
concentration-dependent
manner,
(Eicher
Malki
Tanaka
Yagi-Utsumi
2021).
suggest
increase
gelation
conclusion
cause
well
desiccation-related
IDPs,
thought
preserve
through
prevention
aggregation,
mechanisms
(Chakrabortee
2007;
Koubaa
2019).
conducted
experiments
utilizing
measured
UV-light
(340
nm).
same
making
our
comparable
theirs.
size
exclusion
chromatography
(SEC)
considered
gold
standard
studying
(Hong
2012).
because
2021),
SEC
feasible
will
clog
column
absence
strong
denaturant,
also
perturb
began
mixed
both
before
drying/rehydration.
Dried/rehydrated
samples
showed
minimal
significant
rehydration
relative
hydrated
samples.
show
any
upon
drying-rehydration.
implies
itself.
Moreover,
combination
1g).
Finally,
arithmetic
addition
value
(alone)
compared
(LDH
8)
shows
decrease
dried
state
actual
mixture
1h).
If
would
expect
see
statistically
sum
plus
alone.
observe
unit
components.
that,
helps
1h)
1c).
Using
composed
report
consisting
just
no
significant.
authors,
one-way
ANOVA
Tukey's
post-hoc
1f).
revealed
there
difference
Furthermore,
BSA
indicates
PrCAHS1
(or
inducing)
interpretation
emphasizes
importance
tests
support
conclusions
drawn
experiment,
unfortunately
lacking
(2024).
conclusion,
portions
specifically
helical
linker
region,
previously
shown
enzymes
2023),
evidence
process
lacking.
isolation
Our
data,
alongside
mitigating
drying-induced
dysfunction.
further
like
caution
when
analyzing
IDP
functionality,
important
consider
extending
relevance
observations
partial
native
context
problematic.
A
fragment
take
conformational
differ
its
conformation
Das
2015).
exacerbated
under
environmental
conditions
impart
IDPs
acknowledge
done
hope
commentary
contribute
fruitful
discussion
biology
tardigrades
fascinating
Sourav
Biswas:
Conceptualization;
investigation;
writing
–
original
draft;
methodology;
visualization;
review
editing;
formal
analysis;
curation.
Thomas
C.
Boothby:
editing.
thank
Dr.
Chin-Ju
Park
(Gwangju
Institute
Science
Technology)
providing
raw
associated
manuscript
here.
declare
conflict
interest.
findings
available
corresponding
author
reasonable
request.
Язык: Английский
Protein surface chemistry encodes an adaptive tolerance to desiccation
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Июль 29, 2024
Abstract
Cellular
desiccation
-
the
loss
of
nearly
all
water
from
cell
is
a
recurring
stress
in
an
increasing
number
ecosystems
that
can
drive
protein
unfolding
and
aggregation.
For
cells
to
survive,
at
least
some
proteome
must
resume
function
upon
rehydration.
Which
proteins
tolerate
desiccation,
molecular
determinants
underlie
this
tolerance,
are
largely
unknown.
Here,
we
apply
quantitative
structural
proteomic
mass
spectrometry
show
certain
possess
innate
capacity
rehydration
following
extreme
loss.
Structural
analysis
points
surface
chemistry
as
key
determinant
for
which
test
by
showing
rational
mutants
convert
sensitive
into
tolerant
one.
Desiccation
tolerance
also
has
strong
overlap
with
cellular
function,
highly
responsible
production
small
molecule
building
blocks,
intolerant
involved
energy-consuming
processes
such
ribosome
biogenesis.
As
result,
rehydrated
preferentially
enriched
metabolite
producers
depleted
cell’s
heaviest
consumers.
We
propose
functional
bias
enables
kickstart
their
metabolism
promote
survival
Teaser
Proteins
resist
dryness
tuning
amino
acids
on
surfaces.
Язык: Английский