Crystallization of LEXSY-produced Helminth proteins

LEXSY (Leishmania Expression System) is a eukaryotic protein expression platform based on the S1-classified protozoan L. tarentolae. It combines the advantages of prokaryotic systems (simple handling, fast growth) with those of eukaryotic ones (efficient folding, post-translational modifications).

More than 20 crystal structures of proteins produced with LEXSY have been reported so far, among them human Cu/Zn Superoxide dismutase^[1], Channelrhodopsin 2^[2] or Legumain^[3].

Here LEXSY has been successfully employed by Ebner et al.^[4] to express an active dimeric chitinase from the gastrointestinal nematode Trichuris suis (Pig whipworm), which was suitable for in vitro and in vivo assays as well as for crystallization (Fig. 1).

Figure 1: Three-dimensional structure of the chitinase dimer from T. suis. Proteins have been expressed with LEXSY and crystallized using JBScreen JCSG++. From Ebner et al. (2021)^[4].

JBScreens are designed for efficient and flexible screening of crystallization conditions for soluble proteins, membrane proteins, peptides, nucleic acids, protein-ligand and macromolecular complexes.

• Formulations are systematically arranged in a gradient format
• Chemicals used are of MicroSelect grade for Molecular Biology
• Buffers are prepared as 1 M stock solutions, adjusted to the specific pH value
• Final volume is adjusted with >18 MΩxcm water
• Solutions are sterile-filtered (0.2 μm filter)
• Cacodylate-free
• JBScreen documentation is readily available on the website: Screen Formulation in xml, xls and pdf format, Certificate of analysis (CofA), Material Safety Data Sheet (MSDS)

Crystallization Stock Solutions in larger volumes are available to reproduce previously grown protein crystals.

Crystallization Stock Solution

• High concentrated solution of a precipitant, salt or buffer
• Chemicals are those used in our JBScreens
• Freshly made after receipt of order
• Available in 100 ml volume

Individual JBScreen Condition

• Any condition of a JBScreen with identical chemicals in identical concentrations, e.g. JBScreen JCSG++, condition E2 is 2 M Ammonium sulfate, 100 mM MES pH 6.5 and 200 mM Sodium chloride
• Freshly made after receipt of order
• Available in 10 ml or 100 ml volumes

Need further information?

Please check out the LEXSY section and the crystallography section of our website!

Or contact us by email at: expression@jenabioscience.com and xtals@jenabioscience.com.

References:

[1] Gazdag et al. (2010) Purification and crystallization of human Cu/Zn superoxide dismutase recombinantly produced in the protozoan Leishmania tarentolae. Acta Crystallogr. Sect. F. Struct. Biol. Cryst. Commun. 66:871.
[2] Volkov et al. (2017) Structural insights into ion conduction by channelrhodopsin 2. Science 358, eaan8862.
[3] Zauner et al. (2018) Crystal Structure of Plant Legumain Reveals a Unique Two-Chain State with pH-Dependent Activity Regulation. Plant Cell 30:686.
[4] Ebner et al. (2021) A Helminth-Derived Chitinase Structurally Similar to Mammalian Chitinase Displays Immunomodulatory Properties in Inflammatory Lung Disease. J. Immunol. Res., Article ID 6234836.