Jon Gaffaney Email & Phone Number

United States

Madison, Wisconsin, United States

Madison, Wi

Madison, Wisconsin, United States

University Of Wisconsin - Madison

My responsibilities as a lab manager include but are not limited to budget analysis, consulting on personnel decisions, maintaining current animal and biological safety protocols, and procurement of new equipment and supplies. In addition to these responsibilities, I continue to maintain an active research role in the laboratory continuing to study the biophysics of Doc2 and its regulatory role in SNARE-mediated membrane fusion as well as analyzing experimental data, preparing manuscripts, and training/mentoring graduate students.

Department Of Neuroscience

As a postdoctoral fellow I was responsible for designing and preforming experiments and analyzing data in order to publish our results in highly respected peer-reviewed journals. My studies were focused on characterizing the role of two calcium-sensing proteins (synaptotagmin and Doc2) in regulating synaptic vesicle release. Using a reconstituted membrane fusion system, I characterized the function of recombinant fragments of synaptotagmin, Doc2, or a series of syt/Doc2 chimeras. In addition, I characterized several of their biophysical properties using a variety of techniques including stopped-flow rapid mixing, isothermal calorimetry, liposome co-sedimentation and co-flotation assays. Experiments performed and/or started during this time have resulted in publications in The Journal of Biological Chemistry, Nature Structural and Molecular Biology, Cell, Nature Neuroscience, and Molecular Biology of the Cell.

Grand Forks, North Dakota

As a postdoctoral fellow in my graduate lab I continued my studies on inhibitor and substrate induced conformational changes in the dopamine transporter (DAT). I used the substituted cysteine accessibility method (SCAM) to study the aqueous accessibility of the transporter during inhibitor binding and substrate transport. I generated several cysteine point mutations and developed stable cell lines expressing each of the point mutants. Using these cell lines the accessibility of each cysteine to sulfhydryl reactive compounds following inhibitor or substrate binding was examined. In addition, these cysteine mutants were used to assay DAT function before and after treatment with sulfhydryl reactive compounds.

Grand Forks, North Dakota

As a graduate student I characterized the molecular determinants of the dopamine transporter (DAT) responsible for inhibitor binding and substrate translocation. Prior to the crystallization of the Leucine transporter (LeuT) - a DAT homolog - in 2009 very little structural information was available. To understand how psychoactive molecules such as cocaine and amphetamine interacted with DAT, I utilized a series of cocaine-based photo-activated radioactive labels to native protein from synaptosome isolated from striatal tissue. Using limited proteolysis and epitope specific immunoprecipitation I was able identify regions of DAT that interacted with uptake inhibitors. In addition, I characterized inhibitor induced conformational changes that affected protease sensitivity at specific sites in the protein.

Ph.D., Biochemistry And Molecular Biology

Activities and Societies: Sigma Xi Student Representative - Library committee Student Representative - Faculty meetings Society.

B.S., Biology

Activities and Societies: Medical Post 911; Pre-professional club Intramural sports (Hockey/softball) Work study student under.