Integrated Structural Biology New Core Manager
Change has become a way of life at CBSD’s Integrated Structural Biology Core (ISBC). Last February, we bid farewell to our long-time and multi-talented manager, Dr. Tung-Chung Mou, who accepted a position at Pfizer in Boulder, CO. TC’s accomplishments during his years as manager included the transformation of the “Macromolecular X-ray Diffraction Core” into a fully functional “gene-to-structure” facility – thus, the ISBC –, developing new capabilities in Small Angle X-ray Scattering and, most recently, Cryo-Electron Microscopy. Importantly, TC trained and mentored a long line of students and fellows who conducted their dissertation research and projects with the Core. Dr. Sam Zeng, a recent Ph.D. from Steve Sprang’s lab filled TC’s capacious shoes as interim manager, where, with Cindee Yates-Hansen and Dr. Levi McClelland, he spearheaded the ISBC’s collaboration with the Center of Translational Medicine (CTM) to produce SARS Cov-2 (the virus that causes COVID-19) spike protein to assist the CTM in its vaccine adjuvant research. Last July, Dr. Zeng joined the scientific staff at Ligand pharmaceutical in Oakland CA.
We are pleased to report that, as of January 2021, Dr. Levi McClelland has taken the position of ISBC Manager. Levi earned his Ph.D. in Prof. Bruce Bowler’s lab. There he used stop-flow fluorescence methods to track the folding kinetics of cytochrome c. Levi and his colleagues used X-ray crystallography to describe – for the first time – the catalytic site of cytochrome c lipid peroxidase activity. As a postdoctoral fellow in Dr. Sprang’s Lab, Levi and colleagues determined the crystal structure of the complex of Ric-8A with the G-protein Gai1, a challenging macromolecular complex. Levi is skilled in recombinant protein expression and purification, a variety of protein analytical methods, and X-ray crystallography. Cindee Yates-Hansen will divide her time as a senior technician between the Sprang lab and the ISBC. Cindee is a molecular biologist, skilled in protein expression and purification and X-ray data collection.
The ISBC has completed the construction of its own dedicated laboratory for protein expression and purification, located in ISB-016B. The new lab is conveniently accessible from the X-ray lab where ISBC and Small Molecule X-ray Crystallography Core Facility (Managed by Dr. Dan DeCato) staff and clients grow crystals and collect X-ray diffraction data. The new protein expression lab is equipped with a tissue culture facility for protein expression in eukaryotic cells. Just adjacent to the protein expression and crystallography labs, in ISB-015A, the ISBC houses temperature-controlled incubator shakers for multi-liter scale bacterial growth, medium- and ultra-centrifuges, and a new 80 sq. ft 4° C cold room now under construction. The cold room will allow our staff and collaborators to conduct protein purification procedures under conditions that protect sensitive proteins from denaturation.
If you pass by the ISBC protein expression lab today, you will find that it is in the service of UM’s Covid-19 testing facility, operated by Drs. David Xing and Kim Stanek and a crew of student/staff volunteers. Here Dr. Xing and his colleagues use PCR methods to check for SARS Cov-2 RNA. Please be assured that the no work using intact SARS Cov-2 virus is conducted in ISBC facilities, only processed samples containing RNA.
Our last bit of good news is that ISBC staff and clients will be able to screen and in favorable cases, measure complete X-ray diffraction datasets from protein crystals here at UM. In December 2019, we installed a kappa goniostat that affords a wide range of crystal orientations, and a Pilatus R200K Dectris X-ray detector. Coupled with our MicroMax 007 HF X-ray generator, and Oxford liquid nitrogen crystal cryo-protection system, users will be able to measure high-resolution X-ray datasets from well-diffracting crystals of proteins, protein-protein, protein-ligand, and protein-nucleic acid complexes. As always, the ISBC maintains an ongoing contract with the Stanford Synchrotron Radiation Laboratory (SSRL) for access to high-flux multi-wavelength X-ray beamlines when those facilities are needed to determine crystal structures for ISBC clients. We shall also have access to the Montana State University Cryo-Electron Microscope facility that will be fully operational in the April 2021-June 2021 time frame. The MSU facility under the direction of Prof. Martin Lawrence will house a 200kV Talos Arctica cryo-electron transmission electron microscope and instruments for sample preparation. The ISBC has a 4-node GPU cluster and a full suite of software for EM data processing and macromolecular structure determination.
With new staff, new space, and new equipment, the ISBC is ready to advance the science of UM cell, molecular and biophysical scientists, their students, and staff.