Lipidic cubic phase protein crystallization

Creative Biostructure offers from the single package to the full range of
membrane protein service, starting from DNA cloning to the determination of
protein structure. Collaborating with academic organizations, we have
optimized Lipidic Cubic Phase (LCP) crystallization method to increase the
speed and chance of success.

LCP crystallization is the most commonly used method for membrane protein
structure determination. It has a membrane-mimetic matrix suitable for
stabilization and crystallization of membrane proteins in lipidic environment.
Recent advances in pre-crystallization assays, improvement in crystal imaging,
and commercial availability of LCP instruments greatly attract the attention of
structural biologists. LCP crystallization method has evolved over the years
from using small glass test tubes and mixing lipid with protein solution by
centrifugation to faster and more efficient way of mixing LCP with syringe mixer
and setting up crystallization trials in 96-well plates. Consumption of protein
and lipid per crystallization trial has been reduced by over 100-fold, and the
whole process becomes as easy as setting up vapor diffusion trays.

A gallery of protein and peptide structures obtained by LCP crystallization

Figure 1. A gallery of protein and peptide structures obtained by LCP
crystallization.

To improve the detection efficiency of initial microcrystals in LCP, our scientists

uses trace labeling technology to attach fluorescent dyes to protein molecules.
We also have improved a novel method of protein crystal detection, termed
SONICC (Second Order Nonlinear Imaging of Chiral Crystals), which is devoid
of many drawbacks associated with other imaging techniques. This method
relies on two-photon scattering, which gives virtually no background scattering
from randomly oriented molecules, but produces a strong signal from chiral
molecules arranged in crystals.

Emerging microfluidics approaches capable of handling LCP and other viscous

lipidic mesophases will mature and will likely represent the next major step in
minimizing the consumption of valuable protein material and may allow for
integration of different steps, such as cell-free expression, purification and
crystallization, in a single LCP-lab-on-a-chip. Using this technique, Creative
Biostructure offers the highest-quality crystallographic screening for
membrane proteins with largely increased success rate. We also offer
subsequent crystal optimization, crystal handling and data collection, which
are known to be very difficult in LCP system.
Please feel free to contact us for a detailed quote.

Ordering process

Ordering Process

References

Caffrey M. (2015) “A comprehensive review of the lipid cubic phase or in meso

method for crystallizing membrane and soluble proteins and complexes”. Acta
Cryst F71:3–18.
Cherezov V. (2011) “Lipidic cubic phase technologies for membrane protein
structural studies”. Curr Opin Struct Biol 21(4):559–566.
Landau EM, Rosenbusch JP. (1996) “Lipidic cubic phases: a novel concept for
the crystallization of membrane proteins”. Proc Natl Acad Sci U S
A93(25):14532-14535.