Protein Engineering Core

Division Of Basic Sciences - Nci

Investigators

Linked publications & trials

Abstract

The advent of technological advances in cryo-electron microscopy (cryoEM) has enabled a new avenue for imaging biomolecules, reinvigorating the field of structural biology. Combining cryoEM with the proven techniques of nuclear magnetic resonance (NMR) and X-ray crystallography, several paths open for today's investigator to explore a protein of interest at various scales and in different environments. Each method has strengths and drawbacks, but a unifying factor is a requirement for pure, natively folded protein. Building on our history of providing high-quality purified protein to intramural and extramural collaborators, the Macromolecular Crystallography Laboratory's Protein Engineering Section has been reimagined recently as the Center for Structural Biology's Protein Engineering Core, facilitating the sharing of our expertise with a wider audience. Our competencies include the cloning and expression of pathologically significant proteins in systems that promote high yield and purity. We employ a variety of cloning vectors that convey advantages during expression or structural investigation, producing soluble protein in prokaryotic or eukaryotic expression systems as appropriate. Additionally, the Protein Engineering Core continues to contribute to the development of new and improved methods and reagents for the production of recombinant proteins. In particular, we are interested in understanding how certain highly soluble proteins are able to prevent the aggregation and promote the proper folding of their fusion partners and in exploiting proteases as reagents for the removal of affinity tags. In FY2021, the Protein Engineering Core supported internal and external collaborators by creating expression plasmids, growing production cultures, and performing protein purifications for eight unique projects.