Abstract: A recognized area of mass spectrometry is the analysis of protein conformation and dynamics. One proven approach is to label protein molecules in solution under physiological conditions as the incorporation of the labeling agent is a function of the folded conformation. Hydrogen exchange (HX) methods label the backbone amide hydrogens of proteins with deuterium and the location and magnitude of the labeling can then be determined with mass spectrometry (MS). HX MS studies are particularly well suited for analysis of proteins that will not crystallize, proteins not amenable to NMR, or proteins available in only small quantities. This presentation will explore the rapid growth of HX MS over the past 5 years, discuss the current state of the methodology and address future challenges for practitioners and vendors. Several recent examples of the application of the method will be described, including analysis of proteins of interest to the biopharmaceutical community and analysis of proteins in and at biological membranes.
Biography: John R. Engen is a Professor of Bioanalytical Chemistry at Northeastern University in Boston. Professor Engen holds two B.S. degrees (molecular biology and biochemistry) from Union College and a Ph.D. in Chemistry from the University of Nebraska (working with David L. Smith). He completed postdoctoral work at the European Molecular Biology Laboratory (EMBL) in Heidelberg, Germany and at the Los Alamos National Laboratory. From 2002-2006, he was an Assistant Professor of Chemistry, Biochemistry and Molecular Biology at the University of New Mexico in Albuquerque. In 2012-2013, he was a Visiting Professor of Biochemistry in the laboratory of Prof. F. Ulrich Hartl at the Max Planck Institute of Biochemistry in Martinsried, Germany. Prof. Engen is a Fellow of the European Molecular Biology Organization (EMBO), received the 2009 Arthur F. Findeis Award from the American Chemical Society, and served on the Board of Directors for the American Society for Mass Spectrometry (ASMS) from 2009-2011.
Professor Engen has become a recognized expert in the area of understanding proteins and protein conformation with mass spectrometry. He uses hydrogen-deuterium exchange to probe conformation and dynamics during various activation states. Proteins that are not amenable to mainstream structural techniques such as X-ray diffraction and NMR can be probed with such methods. Such experiments, among other things, can reveal the effects and locations of binding, be diagnostic for proper protein folding, and be used to determine conformational changes during protein function.
Professor Engen has published over 90 papers on the topic of hydrogen exchange in recent years and given hundreds of invited lectures worldwide to academia and industry. He teaches a yearly ASMS short course on protein structural analysis by mass spectrometry. Current research projects in his laboratory include (1) investigations of kinase conformation to understand regulation and aberrant signaling in various disease states including cancer, (2) analysis of the conformation of viral accessory proteins from HIV, (3) studies of protein conformation at biological membranes, and (4) optimization and methods development in hydrogen exchange mass spectrometry.
Prof. Facundo Fernandez (404-385-4432)