GBMSDG Meeting Archives
September 17, 2009
Professor Jonathan V. Sweedler
University of Illinois
Multidimensional Fractionation Strategies
for Quantitative Proteomics
Boston University School of Medicine
Special thanks to Bruker for sponsoring this event. It is the active participation of our sponsors that allows us to keep the fees for membership, dinners, drinks, speakers, etc. so low.
Understanding the functioning of the brain is hampered by a lack of knowledge of the full complement of its signaling molecules, as well as the spatial and temporal interplay of these chemical systems. Mass spectrometry is ideally suited to characterize neuropeptides and hormones, which often become bioactive only after particular post-translational modifications. Several neuropeptidome applications related to cell-cell signaling molecules are highlighted using a variety of high resolution electrospray ionization LC-MS approaches. Using capillary electrophoresis mass spectrometry, the cellular metabolome is measured and related to cell function; as one example, we demonstrate the use of capillary electrophoresis with an ultra-high resolution tandem TOF MS platform for profiling the metabolites and transmitters within individual neurons. Using MALDI-TOF/TOF MS, neuropeptides and hormones are identified directly from tissues including single cells and even individual neuronal processes. Mass spectrometric imaging methods are described that can provide spatial maps of the neuropeptides found in well defined neuronal networks. Using these techniques, multiple neuroactive compounds have been discovered in a range of model organisms ranging from mollusks, insects to vertebrates. Several sampling approaches for mass spectrometry are described that allow the activity dependent release of peptides from select brain regions to be measured. This suite to ultrasensitive mass spectrometry measurement tools allow cell-cell signaling to be followed with unprecedented detail.