Gene-to-Structure Services: Protein Production, Crystallization and Structure Determination

OVERVIEW
Gene-to-Structure services at SARomics Biostructures i
nclude cloning, expression, purification and characterization of a protein using biophysical methods, followed by crystallization and high resolution X-ray structure determination. For the most optimal handling high-throughput protocols developed at the company are used throughout the whole process.

Gene-to-Structure services are performed on a fee-for-service basis and include:

  • Cloning, expression and purification of the protein
  • Biophysical characterization of the protein using dynamic light scattering (DLS), CD spectroscopy, differential scanning fluorimetry (DSF), and if required, NMR spectroscopy.
  • Crystallization of the protein, X-ray data collection and protein structure determination.
  • Protein-ligand co-crystallization and structure determination.
More details on the methods used can be found in our technology pages.

Method summary:

Protein crystal structure determination requires crystals. Our team of crystallographers has many years of experience in protein crystallography and X-ray structure determination, verified, e.g., by the high number of publications co-authored by our team members and company management.
Our integrated protein crystallography gene-to-structure services offer:

  • Protein expression, purification of crystallization-grade protein
  • Crystallization and structure determination.
  • Protein characterization using biophysical methods (dynamic light scattering (DLS), differential scanning fluorometry (DSF), NMR spectroscopy and mass spectrometry).
Our laboratories are well-equipped for protein production and crystallization, for which we use robotic nanoliter-scale screening and optimization of crystallization conditions. For crystallization with ligands we use both soaking of crystals and co-crystallization with the protein target.

X-ray diffraction data are collected at various X-ray sources, primarily European synchrotrons. The structure is normally solved with molecular replacement, however,
de novo structure determination is applied when no experimental structure of the same protein or its homologue is available in the Protein Data Bank. If required, we can also assist in interpretation of the electron density map and analysis of ligand binding. Atomic coordinates of the protein or its complex with ligand, and the X-ray diffraction dataset are retured to clients. If required, we can also assist in the interpretation of the results.