Cell Reports,
Год журнала:
2024,
Номер
43(7), С. 114374 - 114374
Опубликована: Июнь 19, 2024
Morphological
studies
of
skeletal
muscle
tissue
provide
insights
into
the
architecture
fibers,
surrounding
cells,
and
extracellular
matrix
(ECM).
However,
a
spatial
proteomics
analysis
including
muscle-tendon
transition
zone
is
lacking.
Here,
we
prepare
cryotome
sections
mouse
soleus
measure
each
slice
using
short
liquid
chromatography-mass
spectrometry
(LC-MS)
gradients.
We
generate
3,000
high-resolution
protein
profiles
that
serve
as
basis
for
network
to
reveal
complex
junction.
Among
increase
from
tendon,
find
proteins
related
neuronal
activity,
fatty
acid
biosynthesis,
renin-angiotensin
system
(RAS).
Blocking
RAS
in
cultured
tenocytes
losartan
reduces
ECM
synthesis.
Overall,
our
thin
provides
proteome
reveals
acts
an
additional
regulator
within
junctions.
Biology,
Год журнала:
2023,
Номер
12(12), С. 1514 - 1514
Опубликована: Дек. 12, 2023
Immunopeptidomics,
as
the
analysis
of
antigen
peptides
being
presented
to
immune
system
via
major
histocompatibility
complexes
(MHC),
is
seen
an
imperative
tool
for
identifying
epitopes
vaccine
development
treat
cancer
and
viral
bacterial
infections
well
parasites.
The
field
has
made
tremendous
strides
over
last
25
years
but
currently
still
faces
challenges
in
sensitivity
throughput
widespread
applications
personalized
medicine
large
studies.
Cutting-edge
technological
advancements
sample
preparation,
liquid
chromatography
mass
spectrometry,
data
analysis,
however,
are
transforming
field.
This
perspective
showcases
how
advent
single-cell
proteomics
accelerated
this
transformation
immunopeptidomics
recent
will
pave
way
even
more
sensitive
higher-throughput
analyses.
Single
cell
proteomics
by
mass
spectrometry
(SCP)
is
an
emerging
field
of
study
that
has
captured
the
interest
and
imagination
biologists
in
a
wide
array
disciplines.
In
pursuit
this
new
dizzying
technologies
techniques
have
demonstrated
ability
to
quantify
hundreds
few
thousand
proteins
single
mammalian
cells
typical
size.
One
striking
characteristic
these
methods
range
relative
costs
associated
with
analysis
each
cell.
We
attempted
estimate
cost
per
across
17
different
studies
based
on
quotes
we
obtained
for
hardware,
reagents
instrument
support
plans
relation
number
can
be
analyzed
day.
Before
including
labor
or
facilities,
find
analyze
size
from
less
than
<$2
over
$50
The
increase
appears
directly
related
decrease
throughput
as
measured
theoretical
maximum
Perhaps
most
surprising
observation
average
year.
This
when
compared
emergence
RNA
sequencing
where
increased,
cost/cell
decreased
exponentially
first
7
years
field’s
emergence.
While
made
many
assumptions
obtain
estimates,
hope
will
informative
scientists
interested
obtaining
SCP
data
spectrometrists
who
are
considering
entering
field.
provided
spreadsheet
simple
calculator
supplemental
allow
others
adjust
our
calculations
other
variables
which
inevitably
described
future.
Analytical Chemistry,
Год журнала:
2024,
Номер
96(42), С. 16985 - 16993
Опубликована: Окт. 9, 2024
Current
developments
in
single-cell
mass
spectrometry
(MS)
aim
to
deepen
proteome
coverage
while
enhancing
analytical
speed
study
entire
cell
populations,
one
at
a
time.
Custom-built
microanalytical
capillary
electrophoresis
(μCE)
played
critical
role
the
foundation
of
discovery
MS
proteomics.
However,
requirements
for
manual
operation,
substantial
expertise,
and
low
measurement
throughput
have
so
far
hindered
μCE-based
studies
on
large
numbers
cells.
Here,
we
design
construct
robotic
(RoboCap)
platform
that
grants
CE-MS
with
automation
proteomes
limited
less
than
∼100
nL.
RoboCap
remotely
controls
precision
actuators
translate
sample
fused
silica
separation
capillary,
using
vials
this
work.
The
is
hermetically
enclosed
actively
pressurized
inject
∼1-250
nL
into
CE
errors
below
∼5%
relative
standard
deviation
(RSD).
supporting
equipment
were
operated
monitored
custom-written
Virtual
Instrument
(LabView).
Detection
performance
was
validated
empirically
∼5-250
portions
HeLa
digest
trapped
ion
mobility
spectrometer
(timsTOF
PRO).
improved
CE-ESI
utilization
∼20%
from
∼3%
μCE,
closest
reference
technology.
Proof-of-principle
experiments
found
identification
quantification
robustly
return
∼1,800
proteins
(∼13%
RSD)
∼20
ng
earlier-generation
detector.
automates
amounts,
paving
way
electrophoresis-based
high-throughput
Cell Reports,
Год журнала:
2024,
Номер
43(7), С. 114374 - 114374
Опубликована: Июнь 19, 2024
Morphological
studies
of
skeletal
muscle
tissue
provide
insights
into
the
architecture
fibers,
surrounding
cells,
and
extracellular
matrix
(ECM).
However,
a
spatial
proteomics
analysis
including
muscle-tendon
transition
zone
is
lacking.
Here,
we
prepare
cryotome
sections
mouse
soleus
measure
each
slice
using
short
liquid
chromatography-mass
spectrometry
(LC-MS)
gradients.
We
generate
3,000
high-resolution
protein
profiles
that
serve
as
basis
for
network
to
reveal
complex
junction.
Among
increase
from
tendon,
find
proteins
related
neuronal
activity,
fatty
acid
biosynthesis,
renin-angiotensin
system
(RAS).
Blocking
RAS
in
cultured
tenocytes
losartan
reduces
ECM
synthesis.
Overall,
our
thin
provides
proteome
reveals
acts
an
additional
regulator
within
junctions.
