Molecular mechanisms of anesthetics in the nervous system
The major focus of my laboratory is elucidation of the molecular mechanism(s) through which anesthetics act to depress nervous system function. We are specifically interested in identifying the target molecules with which anesthetics interact, and in characterizing these interactions. Two general methods are used in the laboratory to observe the low affinity binding interactions of anesthetics with specific proteins (not observable using conventional binding methodology). 19F-NMR (nuclear magnetic resonance) exchange-spectroscopic methods are used to characterize the kinetics of fluorinated volatile anesthetic binding to purified proteins. To identify the specific nervous system proteins with which anesthetics interact, we have developed photo-labile anesthetic molecules and undertaken photoaffinity labeling studies. Photolabeling studies with fluorinated volatile anesthetics have identified several proteins that contain specific binding sites for the fluorinated volatile anesthetics. More recently, we have developed an anesthetic steroid that can act as a photolabeling reagent. This reagent should allow us to identify the specific CNS proteins that bind anesthetic steroids and to begin a molecular characterization of these sites. The long-term goals of the lab are to synthesize photolabeling reagents for several classes of anesthetics and to use these agents to classify and characterize the binding sites responsible for producing anesthesia.