The major research interest of my laboratory is the correlation of structure and function for membrane protein receptors and channels, primarily neuroreceptors. Utilizing recent biotechnological advances, we successfully overexpressed recombinant human glycine receptors in insect cells at a fairly high level. This receptor is part of a superfamily of ligand-gated ion channels critical in transmission of synaptic signals. Using circular dichroism, and mass spectrometry coupled with chemical modification, we critically examined the topology and structure of this paradigmatic ion channel. In order to continue to characterize the structure of this ion channel, our lab is systematically incorporating cysteine residues at select regions of the receptor and incorporating non-specific crosslinking and cleavage reagents (to map accessibility and proximity using mass spectrometry as a structural tool), fluorescent tags, and spin labels (these latter studies are in collaboration with Dr. Sunil Saxena, Dept. of Chemistry, University of Pittsburgh). We are also engaged in studies examining the effects of lipid composition on the structure and function of the glycine receptor. Crystallographic trials of a soluble form of the extracellular domain of this receptor are also being conducted. Our lab has also initiated collaborative computational studies of the glycine receptor with the laboratories of Dr. Rob Coalson (Dept. of Chemistry, University of Pittsburgh), Dr. Maria Kurnikova (Dept. of Chemistry, Carnegie Mellon University) and Dr. Vasanthi Jayaraman (University of Texas Health Science Center, Houston, Texas). Studies with Dr. Glorioso (past chair of my department at University of Pittsburgh) and a large interdisciplinary team at the University of Pittsburgh seek to develop gene delivery of glycine receptors to peripheral nerves as a way to effectively alleviate pain. Our laboratory also collaborates extensively with faculty both at the University and at other institutions on other projects, some of which are briefly described below. We have recently begun collaborating with an established team of researchers at Duquesne University that are examining the structure and pharmacology of dopamine and serotonin transporters. This team includes Dr. Jeffry Madura, a computational chemist in my department, as well as Drs. David Lapinsky and Chris Surratt in the School of Pharmacy. We have initiated crosslinking studies coupled with mass spectrometric analyses to investigate the binding site of novel photoactivatable drugs that modulate transporter activity.