Equipment Description: The core facility is located in room L.140 of the main campus, 3333 Green Bay Road, North Chicago, IL 60064. The equipment consists of: (1) Rigaku RU-H3R 5.4 kW (fine focus) X-ray generator with Osmic Confocal "Green" optics and a MSC X-Stream 2000 cold jet for crystal cooling. This system also has a CCD video system for mounting crystals. (2) a 3-circle goniometer with a Bruker AXS SMART 6000 CCD detector and an Oxford Cryostream. Computer hardware for data processing include: (1) Dual-CPU (1.2 GHz), 1 GB RAM, 10x80 GB hard disk (0.6 TB) raid array server, running linux; (2) 2 Xeon 153 Dell workstation (2.0 GHz CPU, 1 GB RAM, 120 GB hard drive), running linux; (3) 1 Gateway 500S desktop type (2.0 GHz CPU, 1 GB RAM, 40 GB hard drive), running linux; (4) 1 Custom Monarch desktop (3.20 GHz CPU, 2 GB RAM, 320 GB hard drive with SATA array; and (4) 1 SGI Octane (Irix 6.5). 1*3.0 GHZ, 2.0 GB RAM, 70GB hard drive, Gateway running linux. All data processing computers are maintained and controlled via a master server running NFS in linux. For data collection, computer hardware include :(1) 1 Windows 2000 workstation, running Windows 2000 (Raxis) and (2) 1 Windows XP workstation, running Windows XP (Bruker). All data processing PCs have 3D capable graphics cards and monitors. All machines are NFS / SAMBA / SMB mounted so that data is easily transferable to any pc/server. Backup occurs via raid-array mirroring of disk arrays. All graphics capable machines use 3D IR emitters to allow for hardware enabled stereo viewing with NuVision liquid crystal glasses. Operating Systems: The main Linux distribution is Fedora 3. SGI's run on IRIX 6.5 and the Windows machines on XP and 2000. Software: We are members of the SBGrid community (www.sbgrid.org/) and all software is maintained and operated through their distribution. Printers: HP mono laser, currently only accessible to machines in the X-ray suite via the server. Other hardware: Several low form dewars and two full sets of cryo-tools (tongs, wands, forceps, etc.) are located within the X-ray suite. Liquid nitrogen is stored near the loading facility that adjourns the suite next to the water chiller. Two common storage dewars are available for crystal projects and space is maintained for several dewars owned by individual faculty members.
This equipment serves as the primary in-house structural biology resource for high resolution structure determinations.
The core investigators:
Project
David H. T. Harrison, BMB, Director
Structural Enzymology, Methylglyoxal Synthase, HMG-CoA Synthase, and Hydroxyphenyl pyruvate
Ronald Kaplan, Chair BMB
Mitochondrial transport proteins
D. Eric Walters, BMB
HIV-1 protease/non-peptidyl inhibitors
Kenneth Neet, BMB
p75-NGF complexes
David Mueller, BMB
Yeast ATP synthase
Marc Glucksman, BMB
STRUCTURAL NEUROBIOLOGY of PROCESSING ENZYMES
Jun Choe, BMB
Membrane Protein Structure
Carl Correll, BMB
RNA Chaperones
Adam J. Stein, BMB, Manager
Departmental projects utilizing macromolecular x-ray crystallography
Core Research Interests of the Director and core investigators: Dr Harrison's research uses X-ray crystallography in combination with NMR spectroscopy, Micro Calorimetry, and Mechanistic Enzymology to study the chemical and allosteric mechanisms of methylglyoxal synthase and compare it to the structure and mechanism of the convergently related enzyme triosephosphate isomerase. Additionally, the structures of inhibitor complexes of HMG-CoA Synthase are being determined to design novel anti-cholesterogenic inhibitors in both man and bacteria. Interests of other interdepartmental investigators include: the biochemical mechanisms of enzymes from human pathogens, such as HIV protease, a PPi-dependent PFK from Borrelia, and the surface coat protease from Leishmania. Other research involves the structure determinations of integral membrane proteins such as citrate transport protein, as well as nerve growth factor and its receptor. Other work aims to address problems of energy transduction by studying the FoF1 complex of ATP synthase from yeast. Existing research utilizes a variety of techniques including: directed mutagenesis, kinetic analysis, EPR spectroscopy, surface plasma resonance, CD spectroscopy, DLS, and genetic analysis to elucidate the biological function of macromolecules. The core crystallographic facility greatly complements these methods, and enhances the productivity of the University faculty by allowing researchers to expand their research in new directions. The majority of this work has practical applications to human disease and is funded by the NIH, or other non-profit peer-reviewed organization.
Independent User Time: Users from other institutions (NIH funded or other) are encouragedto apply for time to use the instrumentation (contact Dr.Harrison for details).
Maintenance Information:
Useful X-ray DataProcessing guides: HKL-Manual d*trekwalk through