X-ray and bioNMR Research Core Facility is embedded within the research group of Structural biology. It provides services in X-ray structure determination of small molecules as well as proteins and NMR of biological macromolecules.

Services:

Structure determination of small molecules

Complete servis for determination of small molecules structure is provided. Users bring crystals and receive back structure. Data collection and processing is done at UCT. Result is 3D structure, including stereochemistry, bond lengths, angles and hydrogen bonds. Final structures are deposited in the Cambridge Structural Database (CCDC number). All material (incl. figures) is prepared for publication.

• Contact: Dr. Blanka Klepetářová, office A.01.65, ☎ 340 (klepetarova@uochb.cas.cz).
• Crystal submission form - Acrobat (pdf)   / Word (doc)   
• Crystals: bring crystals in solution in sealed flask to avoid evaporation (crystals should be transparent, no visible imperfections (cracks, twinning), optimal size 0.1-0.5 mm).

Macromolecular X-ray crystallography

We operate crystallization room equipped with crystallization robotics and provide users with training, crystallization plates and basic crystallization screen kits. We organize seminars and workshops on crystallization methods.

• Contact: Dr. Klára Pospíšilová, laboratory A.01.58, ☎ 512 (pospisilova@uochb.cas.cz).
• Shared folder for users on IOCB OwnCloud.

We operate home X-ray diffraction station for macromolecular crystallography.

• Contact: Dr. Jiří Brynda, laboratory A.01.61, ☎ 210 (jiri.brynda@uochb.cas.cz).
• Reservation trough IOCB booking system (for trained users only).

Biomolecular NMR

We operate an NMR instrument (Bruker Avance III™ HD 850 MHz) suitable for protein and nucleic acid studies. We provide training for users both in measurement and data analysis. Simplest measurements of 1D ¹H NMR spectra require 0.16 ml of 50µM protein or short DNA and can provide general information about the molecular fold and presence of unstructured/unfolded regions. These may help to optimize crystallization experiments. Titrations for studies of protein-small molecule interactions using 2D HN correlation spectra are performed routinely by users after short training. Interaction interface on the protein side and Kd can be obtained. The most time-consuming experiments for protein assignment and structure elucidation require 0.35 ml of 200 µM protein. The data collection takes 4 days for protein backbone assignment (necessary for site specific titration interpretation) and additional 10 days for side-chain assignment and structure determination.

• Contact: Dr. Pavel Srb, laboratory A.01.53, ☎ 498 (pavel.srb@uochb.cas.cz).