University of Colorado Cancer Center

Structure determination of macromolecules is performed in collaboration with Shared Resource faculty. Faculty and staff consult with investigators in the project design, protein or nucleic acid expression, purification and sample preparation. The core staff performs data collection and data processing and work with investigators to solve and refine structures.

The Structural Biology Shared Resource enables users to set up initial screens of crystallization conditions using commercially available and custom designed sparse matrix screens manually and with the crystallization robotics systems. Shared Resource faculty members analyze the results of these initial screens to provide guidance in designing or extending the screens to produce crystals for X-ray diffraction studies.

SBSR faculty provide initial data collection on crystals to evaluate project viability. These services provide investigators further directions to improve crystal quality with the goal of a complete structure determination. If high quality crystals are obtained by Shared Resource staff, they collect complete data sets that may be used for structure determination.

The NMR Service collects data and provides interpretation of that data to users studying small molecules such as potential drugs and other therapeutic agents. Data collected includes simple one-dimensional (1D) 1H, 13C and 31P spectra and two-dimensional (2D) COSY, NOESY, ROESY, TOCSY and HSQC correlation spectra.

SBSR staff provide training and consultation in the use of molecular graphics programs to display and analyze molecular structures.

Faculty provide consultation to CU Cancer Center members and other investigators on the appropriate design of experimental conditions for structural analysis. For example this includes design of protein constructs, choice of buffer systems, appropriate methods for data collection. Faculty provide training to PIs, post-doctoral and student researchers in the techniques of sample preparation, data collection, data processing and data interpretation.

NMR spectroscopy is highly suited for screening of intermolecular interactions at atomic resolution. The NMR service of the SBSR collects solution data and provides interpretation of the data to examine interactions between biological macromolecules and other macromolecules or other molecules, including potential drug targets.