Guide to serial synchrotron crystallography
Current Research in Structural Biology,
Год журнала:
2024,
Номер
7, С. 100131 - 100131
Опубликована: Янв. 1, 2024
Serial
crystallography
(SX)
is
an
emerging
technique
that
can
be
used
to
determine
the
noncryogenic
crystal
structure
of
macromolecules
while
minimizing
radiation
damage.
Applying
SX
using
pump-probe
or
mix-and-inject
techniques
enables
observation
time-resolved
molecular
reactions
and
dynamics
in
macromolecules.
After
successful
demonstration
experimental
with
determination
serial
femtosecond
X-ray
free
electron
laser,
this
method
was
adapted
synchrotron,
leading
development
synchrotron
(SSX).
SSX
offers
new
opportunities
for
researchers
leverage
techniques,
contributing
advancement
structural
biology
offering
a
deeper
understanding
function
This
review
covers
background
advantages
its
approach.
It
also
discusses
important
considerations
when
conducting
experiments.
Язык: Английский
Impact of Diffraction Data Volume on Data Quality in Serial Crystallography
Crystals,
Год журнала:
2025,
Номер
15(2), С. 104 - 104
Опубликована: Янв. 21, 2025
Serial
crystallography
(SX)
enables
macromolecular
structure
determination
at
biologically
relevant
temperatures
while
minimizing
radiation
damage.
This
technique
relies
on
processing
numerous
diffraction
images
from
multiple
crystals
to
construct
a
complete
dataset
for
three-dimensional
determination.
Although
increasing
the
volume
of
SX
data
improves
quality,
excessive
collection
reduces
beamtime
efficiency.
Therefore,
understanding
relationship
between
and
quality
is
crucial
efficient
use
beamtime.
In
this
study,
serial
synchrotron
datasets
lysozyme
glucose
isomerase
were
analyzed
assess
impact
varying
volumes
statistics
structural
outcomes.
Data
refinement
metrics
improved
as
integrated
increased;
however,
rate
improvement
in
was
not
proportional
number
patterns.
Furthermore,
decreased
beyond
certain
threshold
volume.
These
findings
expand
our
provide
insights
into
optimizing
efficiency
processing.
Язык: Английский
Comparative Analysis of Room Temperature Structures Determined by Macromolecular and Serial Crystallography
Crystals,
Год журнала:
2024,
Номер
14(3), С. 276 - 276
Опубликована: Март 14, 2024
Temperature
directly
influences
the
function
and
structure
of
proteins.
Crystal
structures
determined
at
room
temperature
offer
more
biologically
relevant
structural
information
regarding
flexibility,
rigidity,
thermal
motion
than
those
by
conventional
cryocrystallography.
can
be
using
macromolecular
crystallography
(MX)
or
serial
(SX)
techniques.
Among
these,
MX
may
theoretically
affected
radiation
damage
X-ray
heating,
potentially
resulting
in
differences
between
SX,
but
this
has
not
been
fully
elucidated.
In
study,
xylanase
GH11
from
Thermoanaerobacterium
saccharolyticum
was
(RT-TsaGH11-MX).
The
RT-TsaGH11-MX
exhibited
both
open
closed
conformations
substrate-binding
cleft
within
β-sandwich
fold.
showed
distinct
changes
molecular
flexibility
when
compared
with
RT-TsaGH11
via
synchrotron
crystallography.
notable
conformation
induced
heating.
These
findings
will
broaden
our
understanding
potential
limitations
MX.
Язык: Английский
Real-time observation of a metal complex-driven reaction intermediate using a porous protein crystal and serial femtosecond crystallography
Nature Communications,
Год журнала:
2024,
Номер
15(1)
Опубликована: Июнь 29, 2024
Abstract
Determining
short-lived
intermediate
structures
in
chemical
reactions
is
challenging.
Although
ultrafast
spectroscopic
methods
can
detect
the
formation
of
transient
intermediates,
real-space
cannot
be
determined
directly
from
such
studies.
Time-resolved
serial
femtosecond
crystallography
(TR-SFX)
has
recently
proven
to
a
powerful
method
for
capturing
molecular
changes
proteins
on
timescales.
However,
methodology
been
mostly
applied
natural
proteins/enzymes
and
limited
promoted
by
synthetic
molecules
due
structure
determination
challenges.
This
work
demonstrates
applicability
TR-SFX
investigations
reaction
mechanisms
metal
complexes.
We
fix
light-induced
CO-releasing
Mn(CO)
3
center
porous
hen
egg
white
lysozyme
(HEWL)
microcrystals.
By
controlling
light
exposure
time,
we
capture
real-time
Mn-carbonyl
intermediates
during
CO
release
reaction.
The
asymmetric
protein
environment
found
influence
order
release.
experimentally-observed
path
agrees
with
quantum
mechanical
calculations.
Therefore,
our
demonstration
offers
new
approach
visualize
atomic-level
small
using
determination.
advance
holds
potential
facilitate
design
artificial
metalloenzymes
precise
mechanisms,
empowering
design,
control
development
innovative
reactions.
Язык: Английский
Effects of Beam Center Position Shifts on Data Processing in Serial Crystallography
Crystals,
Год журнала:
2025,
Номер
15(2), С. 185 - 185
Опубликована: Фев. 15, 2025
Serial
crystallography
(SX)
enables
the
determination
of
biologically
relevant
structures
at
room
temperature
while
minimizing
radiation
damage.
During
SX
experiments,
beam
center
on
diffraction
images
can
shift
due
to
X-ray
movements
or
detector
displacement.
Consequently,
geometry
file
for
is
optimized;
however,
effects
deviations
from
optimal
position
data
processing
efficiency
remain
unclear.
This
study
examines
how
changes
in
influence
quality
by
analyzing
indexing
and
structure
refinement
lysozyme
glucose
isomerase
datasets,
considering
shifts
parameter.
The
results
revealed
that
as
deviated
farther
its
position,
declined,
with
extent
effect
varying
significantly
across
algorithms.
XDS
MOSFLM
algorithms
maintained
high
efficiencies
(>90%)
≤4
pixels
(688
μm)
≤2
(344
μm),
respectively,
compared
processed
optimized
center.
Conversely,
DirAx
XGANDALF
exhibited
below
90%
a
two-pixel
These
findings
enhance
our
understanding
affect
provide
valuable
insights
developing
effective
strategies.
Язык: Английский
Application of Fixed-Target Microcrystal Delivery Systems for Serial Femtosecond Crystallography at PAL-XFEL
Analytica—A Journal of Analytical Chemistry and Chemical Analysis,
Год журнала:
2025,
Номер
6(1), С. 7 - 7
Опубликована: Фев. 27, 2025
Serial
femtosecond
crystallography
(SFX)
using
X-ray
free-electron
lasers
(XFELs)
enables
the
determination
of
biological
and
chemical
structures
without
radiation
damage.
In
SFX
experiments,
a
sample
delivery
system
is
essential
for
delivering
numerous
crystals
to
interaction
point
in
serial
stable
manner.
Among
various
methods,
fixed-target
(FT)
straightforward
widely
used
collecting
data
due
its
advantages
low
consumption
reduced
physical
damage
during
collection.
Here,
we
review
development
FT
with
Pohang
Accelerator
Laboratory
laser
(PAL-XFEL).
The
specifications
operational
conditions
FT-SFX
chamber
are
described.
design,
specifications,
applications
one-
two-dimensional
holders
developed
PAL-XFEL
also
detailed.
Furthermore,
each
discussed.
This
not
only
provides
valuable
information
on
experiments
but
offers
insights
into
systems.
Язык: Английский
Minimized Sample Consumption for Time-Resolved Serial Crystallography Applied to the Redox Cycle of Human NQO1
bioRxiv (Cold Spring Harbor Laboratory),
Год журнала:
2024,
Номер
unknown
Опубликована: Апрель 29, 2024
Abstract
Sample
consumption
for
serial
femtosecond
crystallography
(SFX)
with
X-ray
free
electron
lasers
(XFELs)
remains
a
major
limitation
preventing
broader
use
of
this
powerful
technology
in
macromolecular
crystallography.
This
drawback
is
exacerbated
the
case
time-resolved
(TR)-SFX
experiments,
where
amount
sample
required
per
reaction
time
point
multiplied
by
number
points
investigated.
Thus,
order
to
reduce
consumption,
here
we
demonstrate
implementation
segmented
droplet
generation
conjunction
mix-and-inject
approach
TR
studies
on
NAD(P)H:quinone
oxidoreductase
1
(NQO1).
We
present
design
and
application
injectors
Single
Particles,
Clusters,
Biomolecules
&
Serial
Femtosecond
Crystallography
(SPB/SFX)
instrument
at
European
XFEL
(EuXFEL)
synchronized
injection
that
allows
liquid
phase
protein
crystal
injection.
carried
out
TR-crystallography
experiments
305
ms
1190
NQO1
its
coenzyme
NADH.
With
successful
TR-SFX
approach,
up
97%
has
been
conserved
compared
continuous
suspension
gas
dynamic
virtual
nozzle.
Furthermore,
obtained
structural
information
NADH
an
important
part
future
elucidation
mechanism
crucial
therapeutic
enzyme.
Язык: Английский
Application of Serial Crystallography for Merging Incomplete Macromolecular Crystallography Datasets
Crystals,
Год журнала:
2024,
Номер
14(12), С. 1012 - 1012
Опубликована: Ноя. 22, 2024
In
macromolecular
crystallography
(MX),
a
complete
diffraction
dataset
is
essential
for
determining
the
three-dimensional
structure.
However,
collecting
experimental
using
single
crystal
frequently
unsuccessful
due
to
poor
quality
or
radiation
damage,
resulting
in
collection
of
multiple
incomplete
datasets.
This
issue
can
be
solved
by
merging
datasets
generate
dataset.
study
introduced
new
approach
from
MX
serial
(SX).
Six
β-glucosidase
Thermoanaerobacterium
saccharolyticum
(TsaBgl)
were
processed
CrystFEL,
an
SX
program.
The
statistics
merged
data,
such
as
completeness,
CC,
CC*,
Rsplit,
Rwork,
and
Rfree,
demonstrated
dataset,
indicating
improved
compared
with
enabling
structural
determination.
Also,
was
four
different
indexing
algorithms,
their
compared.
conclusion,
this
generating
will
provide
opportunity
structure
macromolecules
Язык: Английский