bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2023,
Номер
unknown
Опубликована: Окт. 20, 2023
The
ability
to
sequence
single
protein
molecules
in
their
native,
full-length
form
would
enable
a
more
comprehensive
understanding
of
proteomic
diversity.
Current
technologies,
however,
are
limited
achieving
this
goal.
Here,
we
establish
method
for
long-range,
single-molecule
reading
intact
strands
on
commercial
nanopore
sensor
array.
By
using
the
ClpX
unfoldase
ratchet
proteins
through
CsgG
nanopore,
achieve
single-amino
acid
level
sensitivity,
enabling
sequencing
combinations
amino
substitutions
across
long
strands.
For
greater
accuracy,
demonstrate
reread
individual
molecules,
spanning
hundreds
acids
length,
multiple
times,
and
explore
potential
high
accuracy
barcode
sequencing.
Further,
develop
biophysical
model
that
can
simulate
raw
signals
ACS Nano,
Год журнала:
2024,
Номер
18(36), С. 25155 - 25169
Опубликована: Авг. 27, 2024
Steviol
glycosides
(SGs)
are
a
class
of
high-potency
noncalorie
natural
sweeteners
made
up
common
diterpenoid
core
and
varying
glycans.
Thus,
the
diversity
glycans
in
composition,
linkage,
isomerism
results
tremendous
structural
complexity
SG
family,
which
poses
challenges
for
precise
identification
leads
to
fact
that
SGs
frequently
used
mixtures
their
variances
biological
activity
remain
largely
unexplored.
Here
we
show
wild-type
aerolysin
nanopore
can
detect
discriminate
diverse
species
through
modulable
electro-osmotic
flow
effect
at
varied
applied
voltages.
At
low
voltages,
neutral
molecule
was
drawn
stuck
pore
entrance
due
an
energy
barrier
around
R220
sites.
The
ensuing
binding
events
enable
majority
species.
Increasing
voltage
break
cause
translocation
events,
allowing
unambiguous
several
pairs
differing
by
only
one
hydroxyl
group
recognition
accumulation
from
multiple
sensing
regions
Based
on
data
15
SGs,
deep
learning-based
artificial
intelligence
(AI)
model
created
process
individual
blockage
achieving
rapid,
automated,
single-molecule
quantification
real
samples.
This
work
highlights
value
analysis
complex
glycans-containing
glycosides,
as
well
potential
sensitive
rapid
quality
assurance
glycoside
products
with
use
AI.
Proceedings of the National Academy of Sciences,
Год журнала:
2024,
Номер
121(38)
Опубликована: Сен. 12, 2024
The
transport
of
biopolymers
across
nanopores
is
an
important
biological
process
currently
under
investigation
for
the
rapid
analysis
DNA
and
proteins.
While
generally
understood,
methods
to
induce
unfolded
protein
translocation
have
only
recently
been
discovered
(Yu
et
al.,
2023,
Sauciuc
2023).
Here,
we
found
that
during
electroosmotically
driven
polypeptides,
blob-like
structures
typically
form
inside
nanopores,
often
obstructing
their
preventing
addressing
individual
amino
acids.
This
in
contrast
with
electrophoretic
DNA,
where
formation
such
has
not
reported.
Comparisons
between
different
nanopore
sizes
shapes
modifications
by
surface
chemistries
allowed
formulating
a
mechanism
blob
formation.
We
also
show
single-file
can
be
achieved
using
1)
entry
internal
diameter
smaller
than
persistence
length
polymer,
2)
nonsticky
(i.e
.,
nonaromatic)
inner
surface,
3)
moderate
velocities.
These
experiments
provide
basis
understanding
polypeptide
confinement
improving
design
engineering
analysis.
Molecular
dynamics
shows
that
single
amino
acids
can
be
distinguished
via
translocation
through
a
graphene
nanopore.
We
probe
the
effect
of
their
orientation
on
current
blockade.
In
the
decades
following
discovery
that
genes
encode
proteins,
scientists
have
tried
to
exhaustively
and
comprehensively
characterize
human
genome.
Recent
advances
in
computational
methods
along
with
transcriptomic
proteomic
techniques
now
shown
historically
non-coding
genomic
regions
may
contain
non-canonical
open
reading
frames
(ncORFs),
which
functional
miniproteins
or
otherwise
exert
regulatory
activity
through
coding-independent
functions.
Increasingly,
it
is
clear
these
ncORFs
play
critical
roles
major
diseases
such
as
cancer.
this
review,
we
summarize
history
current
progress
of
ncORF
research
explore
known
functions
they
encode.
We
particularly
highlight
emerging
body
evidence
supporting
a
role
for
contributions
Finally,
provide
blueprint
high-priority
areas
future
cancer,
focusing
on
detection,
characterization,
therapeutic
intervention.
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2023,
Номер
unknown
Опубликована: Окт. 20, 2023
The
ability
to
sequence
single
protein
molecules
in
their
native,
full-length
form
would
enable
a
more
comprehensive
understanding
of
proteomic
diversity.
Current
technologies,
however,
are
limited
achieving
this
goal.
Here,
we
establish
method
for
long-range,
single-molecule
reading
intact
strands
on
commercial
nanopore
sensor
array.
By
using
the
ClpX
unfoldase
ratchet
proteins
through
CsgG
nanopore,
achieve
single-amino
acid
level
sensitivity,
enabling
sequencing
combinations
amino
substitutions
across
long
strands.
For
greater
accuracy,
demonstrate
reread
individual
molecules,
spanning
hundreds
acids
length,
multiple
times,
and
explore
potential
high
accuracy
barcode
sequencing.
Further,
develop
biophysical
model
that
can
simulate
raw
signals
