The Structural Biology Core provides UCCC researchers access to instrumentation and expertise in the use of X-ray Crystallography and Nuclear Magnetic Resonance Spectroscopy. The Core aims to promote structural studies in the molecular basis of cellular processes that regulate the development and progression of cancer. We enable member access to crystallization facilities, X-ray crystallography facilities, and Synchrotron X-ray data collection.
Services Offered
Structure Determination of Macromolecules (Proteins/Nucleic Acids)
Structure determination of macromolecules is performed in collaboration with Core 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.
Preliminary Crystallization Screening
The core faculty provides initial screens of crystallization conditions using commercially available and custom designed sparse matrix screens. Core 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.
Preliminary Crystal Screening and Data Collection
X-ray Crystal Structure of LUSH-cVA complex.
The VA is completely enveloped within the protein
Core 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 Core staff collect complete data sets that may be used for structure determination.
Solution structural analysis of small molecules
The NMR Core 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.
Molecular Visualization
Core staff provide training and consultation in the use of molecular graphics programs to display and analyze molecular structures. The Structural Biology Core has 3 Silicon Graphics computers equipped with three-dimensional viewing hardware and software for this purpose.
Experimental Design and Training
Faculty provide consultation to 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.
Equipment
X-ray Instrumentation
Rigaku/MSC RUH3R x-ray generator equipped with confocal optics
Varian INOVA 500 MHz Spectrometer
RAXIS IV ++ x-ray area detector
Rigaku/MSC X-STREAM cryo-cooling apparatus
RAXIS IV++ area detector
Confocal optics
MSC X-STREAM cryo-cooling apparatus
NMR Instrumentation
Varian INOVA 500 MHz system
5 mm triple resonance 1H/13C/15N probe with Z-axis gradient with sensitivity of 850:1 (ETB)
3 mm triple resonance 1H/13C/15N probe with Z-axis gradient with ETB sensitivity of 850:1
Varian INOVA 600 MHz system
5 mm triple resonance 1H/13C/15N probe with Z-axis gradient
5mm triple resonance 1H/13C/15N cryogenically cooled probe with Z-axis gradients
Varian INOVA 800 MHz system
5 mm triple resonance 1H/13C/15N probe with Z-axis gradient
Varian 900 MHz Direct Drive NMR system
5 mm triple resonance 1H/13C/15N probe with Z-axis gradient
Structural Biology CoreLocations
500, 600 MHz and 900 MHz NMR Spectrometers
Anschutz Medical Campus
RC1-South, Room 1300
800 MHz NMR spectrometer
CU Boulder Campus
Center for Innovation & Creativity, Room 193A
X-Ray Crystallography Core
Anschutz Medical Campus
RC1-South, Room 1301
Leadership & Staff
David Jones, PhD
NMR Director
david.jones@ucdenver.edu
303-724-3600
Mair Churchill, PhD
X-Ray Crystallography Director
mair.churchill@ucdenver.edu
303-724-3670
Rui Zhao, PhD
X-Ray Crystallography Manager
rui.zhao@ucdenver.edu
303-724-3269
©2009 University of Colorado Cancer Center
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