Small Angle X-Ray

Scattering
SAXS
Information about;
macromolecular folding,
unfolding,
aggregation,
extended conformations,
flexibly linked domains,
shape,
conformation
assembly state in solution

Resolution range : 10 Å to 50 Å
• Averaged particle sizes, shapes,
distrubition, surface to volume
ratio
• Material can be solid/liquid or
they can contain solid, liquid or
gaseous domains/particles
• Can be apply on;
• Colloids of all types,
• Metals,
• Cement,
• Oil,
• Polymers,
• Plastics,
• Proteins,
• Food,
• Pharmaceuticals
• Before scattering X-rays are transformed into a well-
defined line-shaped or point-shaped beam. This process is
called collimation. Each collimation type in a SAXS
system is ideal for different applications.
• A line-collimated beam has the advantage that it combines
a high photon flux with a high scattering volume – which
means measurement times can be dramatically shorter than
with point collimation. The drawback of a line-collimated
beam is that it can only probe isotropic samples.
Therefore, a line-shaped beam is preferable for analyzing
weakly scattering samples, such as proteins and other soft
matter.
• A point-collimated beam can be used to also analyze
anisotropic samples, such as fibers or porous solids. Point
collimation allows you to probe small sample areas and
determine their local nanostructure, with the drawback of
longer measuring times.
• SAXS and X-ray crystallography are fundamentally similar techniques
and can share most of the hardware required to generate, prepare, and
detect X-rays.

• SAXS data can be used to determine the low-resolution structures of

macromolecules without any additional experimental information.

• Information derived from SAXS data can be useful both prior to and
after high-resolution structures are solved
• Fundamental difference between solution scattering and X-ray
crystallography lies in the relative organization of target molecules during
data collection.

• Solution scattering; the signal from all orientations of the target molecules, relative
to one another and the experimental apparatus, are averaged together.

• X-ray crystallography the molecules are highly organized within a crystal lattice.
Diffraction from a crystal lattice gives rise to discrete diffraction maxima that are
caused by the convolution of the crystal lattice onto the continuous transform due
to the atomic positions and provides enormously greater signal.

• In crystal lattice the orientation of the molecules allows the diffraction data to
retain information about atomic positions in three-dimensional space, which is lost
in SAXS data collected on molecules in solution that are orientationally averaged.
• Ab-initio shape determination can be applied to reconstruct the low-resolution envelopes of the
protein if atomic structures are not available.
• DAMMIN, DALAI_GA, SAXS3D, or GA_STRUCT can be used.

• If the number of residues of the protein is known,

• GASBOR has the advantages of providing significantly more sophisticated penalties that con-strain the models
and of not prohibiting the generation of cavities and other anisotropic shapes

• Comparison of multiple reconstructions is extremely important to verify the stability of the

solution. Multiple repetition of the modeling process significantly decreases the risk of inferring
erroneous shapes.
• DAMMAVER aligns, averages, checks the uniformity, and computes a probability map of the given ab-initio
models.

• If the atomic structure of the sample is known or an atomic model has been proposed,
comparison of theoretical SAXS profile with the experimental data is the first step in the
structure evaluation.
• CRYSOL

• Ab-initio shapes and atomic resolution models may be superimposed by using;

• SUPCOMB or SITUS.
• Disagreement between atomic resolution models and SAXS results;

• The presence of heterogeneous multimerization. This may be apparent if the

molecular weight is determined.
• OLIGOMER can take the calculated scattering profiles of several proposed multimers and
determine the fractional concentration of each multimer required to best fit the data. An
excellent fit with OLIGOMER and a poor fit with CRYSOL implies heterogeneous
multimerization.

• Flexibility and truncated loops (common in crystal structures where disordered

regions are not built into models).
• Generating a variety of conformations of the flexible regions and using OLIGOMER may
improve the final fit.