This core facility will support the research of the COBRE investigators. Specifically, high resolution LC/MS/MS instrumentation housed in the facility will be used in Project 2 (Porter) for identification of proinflammatory arachidonic acid metabolites generated by CGRP stimulated THP-1 cells, and to quantitate the amount of prostaglandin compounds hypothesized to be produced from an increased expression of COX-2. These measurements will benefit studies of the characterization of the neuroinflammatory CGRP receptor. In Project 3 (Picklo), MS will be utilized in studies of the mechanism of HNE metabolism in CNS tissue. High resolution LC/MS will be used to identify and quantitate HNE metabolites, and MALDI/TOF will be used to identify novel detoxification enzymes. Project 4 (Murphy) will utilize the mass spectrometry core facility to fulfill analysis requirements for two specific aims.
These aims address phospholipid head group flux and turnover in neuronal, astroglial, and stably transfected HEK-293 cell lines. LC/MS and GC/MS will be used to measure labeled and unlabeled phospholipids to study the kinetics of compound incorporation. The core facility will consist of three instruments. One instrument will be a high-resolution tandem mass spectrometer (MS/MS, QQTOF geometry) with an ESI source and a liquid chromatography interface. In addition, we will acquire a MALDI/Time of Flight (TOF) instrument and a gas chromatography 1 ion trap system. The combination of these instruments will result in a facility capable of providing molecular weight and structural information for all of the wide variety of samples that will be generated in this COBRE. The facility responsibilities (training users and helping with data analysis, operation and maintenance, assuring that the standard operating procedures are followed) will be divided between between the facility director and a research technician.
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