The depth and breadth of the protein biochemistry expertise among the PCHPI membership is outstanding and contributes to the center's successes in several areas. The PCHPI Protein Core is an anchor for most of the structure determination projects, as it develops novel protein expression protocols, has the resources to produce high quality protein in the large amounts required for structural methodologies, and plays a key role in validating structural observations by mutagenesis assays and other means. The expertise of Dr. Jinwoo Ahn, the director of protein core, is complemented by several PCHPI collaborators including, but not limited to, Drs. Chris Aiken, Alan Engelman, Peter Prevelige, and Jacek Skowronski.
Protein Production & Biochemical Characterization (Protein Core)
Director: Jinwoo Ahn, PhD
The amounts and properties of purified proteins that are necessary for NMR or crystallographic structure determination are frequently underestimated when planning structural biology projects and, thus, constitute a major bottleneck for structural work. The Protein Production core places strong emphasis on providing high quality material for further analysis. Production of correctly folded and mono-dispersed protein solutions is particularly important for structural work.
The Protein Production Core is primarily responsible for producing milligram quantities of homogenous, pure (>95%), virus encoded proteins, human cellular partners, and their complexes for structural studies. Secondarily, the core supports biophysical and bioanalytical characterization of these. The PCHPI makes use of a protein production suite in the Department of Structural Biology that supports protein expression and purification from different organisms, ranging from bacteria to mammalian cells. The facility has four separate rooms and necessary instrumentation to express protein in bacteria, yeast, insect cells, and mammalian cells. For high resolution top-down and bottom-up proteomics work, we have access to an FT MS with a 12 Tesla magnet , which allow molecular weight determination and sequencing of polypeptide mixtures with highest accuracy.