The Laboratory of Biomolecular Structure and Function (LBSF) was created in 2015 to serve scientists of Oklahoma City, in particular OUHSC and OMRF, that are interested in structural aspects of their macromolecular targets. However, the LBSF is also open to academic and industrial users from inside and outside Oklahoma. The LBSF is part of the OUHSC Vice President Research Core Facilities. The LBSF was created based on the Macromolecular Crystallography Laboratory of the Department of Biochemistry and Molecular Biology. This earlier lab was created in 1998 and became a core facility for the Oklahoma Center of Biomedical Research Excellence (COBRE) in Structural Biology (OCSB) in 2013. The LBSF is one of two sites that make up the Biomolecular Structure Core of the OCSB. The other site is the Macromolecular Crystallography Laboratory (MCL) at OU-Norman. Users of the LBSF also have access to the MCL, which has a complementary set of instruments and expertise.
The Protein Expression and Purification (PEP) facility is associated with the LBSF and is set up to help users scale up protein expression to the multi-milligram level for structural, biophysical, and other studies. The PEP has a temperature-controlled shaker for inducing protein expression at low temperatures to improve protein solubility. It also has a ATKA chromatography system and preparative scale size exclusion columns.
The LBSF has a 2014 Wyatt plate reading dynamics light scattering
instrument that enables high-throughput data collection with no user
intervention for evaluating the oligomerization states of proteins.
Monodisperse solutions with 5-20 mg/ml of protein are required for
crystallization experiments, small angle x-ray scattering experiments,
and cryo-electron microscopy (cryo-EM) single-particle analysis experiments.
Roughly 200 microliters of 95% pure, monodisperse protein sample at a concentration
of 5-20 mg/ml are needed to set up ten 96-well trays with the 2018 Art
Robbins Gryphon LCP crystallization robot. The robot can also perform
crystallization experiments in the lipidic cubic phase to crystallize membrane proteins.
The LBSF is equipped with modern crystallographic equipment and computers: A 2015 Rigaku 007 generator, R-axis IV and Mar345 image plate detectors, Osmic mirrors on both ports, Rigaku and Oxford Systems crystal cooling systems. The R-AXIS IV has an optional helium path for working with large unit cells.
The D-Trek, XDS, Mosflm, and HKL2000 software packages are available for X-ray diffraction data processing. Any of the Phenix, CCP4, and CNS program suites can be used for structure determination and refinement, while COOT, Chimera and PyMOL are offered for graphical representation of the results.
The LBSF has a 2019 Leica cryo-plunger to prepare cryo-grids for cryo-EM work at national centers.
Help with grants
Dr. Mooers helps users with the writing of grants that include structural studies.
Services and Fees, Internal and [External]
- Molecular cloning, 22.07/h [$43.96/h]
- Protein over-expression, purification, or both; 22.07 [$43.96]
- 96-well crystallization tray with robot for finding initial lead, $29.43/tray [$40.23/tray]
- 24-well crystallization tray for crystal size optimization, $34.61/tray [$67.32/tray]
- 15-well crystallization tray for crystal size optimization, $34.04/tray [$68.81/tray]
- Crystal handling services, $22.07/h [$43.96/h]
- evaluating results of crystallization experiments
- screening of crystals for diffraction at room temperature
- screening of crystals for cryo-conditions
- local X-ray diffraction data collection
- X-ray computation services, 22.07/h [$43.96/h]
- Remote data collection at synchrotron radiation lab
- X-ray diffraction data processing
- Structure determination
- Structure refinement
- Structure validation analysis
- Structure deposition in the Protein Data Bank
- Figure making for publication
- Molecular modeling services by Dr. Mather, $75.00/h [$75.00/h]
Main Users
- Blaine H. M. Mooers, PhD
- Marie Hanigan, PhD
- Augen Pioszak, PhD
- Franklin A. Hays, PhD
- Xin Zhang, MD, PhD
- Jie Wu, PhD
- Ralf Janknecht, PhD
- Wei-Qun Ding, PhD
- Jody Summers, PhD
- Jialing Lin, PhD
- Ian Dunn, MD
Staff
- To-be-named, PhD – Staff Scientist (BRC 406 and 472)
- Tim Mather, PhD - Molecular Modeling Facility (BRC 238)
Location
The Laboratory is located in the Biomedical Research Center Building on the University of Oklahoma Health Sciences Center. The 975 NE 10th Street Biomedical Research Center Building Oklahoma City, Oklahoma 73104 (405) 271-8300 Email Contact
Download a Campus Map
Recent crystal structures determined by the LBSF
Click to view interactive PDB files
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RET kinase structures:
- 7JU6 RET protein tyrosine kinase with selpercatinib
Useful websites
- Biologist's guide to using the OU supercomputer: slides video
- Macromolecular Crystallography Laboratory, OU-Norman Campus
- SBGrid Consortium
- American Crystallography Association
- International Union of Crystallography
- HKL-2000 manual
- Rigaku’s homepage
- Mar345 Manual
- Crystal Clear manual (includes d*TREK)
- XDS Manual
- Phenix manual
- CCP4 homepage
- CNS manual
- SHELX homepage
- Graphics program COOT manual
- Biological Small Angle Scattering Links
- Molecular Graphics Links from the 'PyMOL' course
- PyMOL for molecular artwork; download site. First-year GPiBS students have the incentive version of PyMOL. We are still trying to get a site license for the University.
- Open-source PyMOL for Windows, open source is functional but exporting movies takes longer.
- Open-source PyMOL for Mac, the macports version installs works well.
- EasyPyMOL, get started for horizontal scripting in PyMOL
- pymolshortcuts, shortcuts for greater productivity in PyMOL
- pymolsnips, PyMOL snippet library for text editors
- pymolpysnips, PyMOL snippet library for Jupyter Notebooks
- UCSF Chimera, useful for making images and structure analysis, especially with cryo-EM maps
- Jmol, a molecular graphics program based on Java.
- Ligplot, a program for creation of ligand-protein interaction diagrams
- Ligplot manual
- More crystallographic links
Upcoming events
- A 5-week graduate course on the Methods of Structural Biology (course director: Dr. Pioszak) in the first session, Spring Semester 2023.
- A 5-week graduate course on Biochemical and Biophysical Instrumentation (course director Dr. Lin) includes two lectures on X-ray diffraction experiments, third session, Spring Semester 2023.
- 2022 SSRL/LCLS Users Meeting was virtual for the third year. The videos of the talks are available online for the past three years if you registered for the meeting. This meeting attracts 1600 people from around the world. Attendance of the virtual meetings has been free. Dr. Mooers has co-chaired a workshop on computational methods in the structural sciences for the past three years and plans to do so again in 2023. Click for more information.