AVB Training Program: Biochemistry/Biophysics
Students and postdoctoral fellows have the opportunity to participate in a variety of projects ranging from the determination of protein primary structure to the study of enzyme mechanisms. Participants receive training in the use of routine biochemical procedures including ultracentrifugal separation of lipoproteins, radiolabeling of proteins and lipids, electrophoresis, and standard assays for the determination of protein and lipid. In addition, trainees are taught to use more sophisticated antibody procedures such ELISAs, Radioimmunoassays, and Western blotting techniques. Participants may receive training in protein sequencing and amino acid analysis using state of the art technology. In addition, training in the determination of carbohydrate and lipid content using HPLC and GC is also available. Instruction in the design and synthesis of peptides as models for apolipoproteins, for membrane association and/or lysis, or for targeting cellular receptors are provided. Training in the use of circular dichroic spectropolarimetry for the determination of protein secondary structure is also offered. Lipid and/or protein transfer between surfaces may be examined by rapid reaction techniques which combine a stopped flow apparatus with fluorescence or UV/Visible spectroscopy. Participants may also be trained in the use of fluorescent probes to examine parameters such as membrane polarity and fluidity. Instruction in the application of kinetic analysis and fluorescence theory to practical problems accompanies the hands-on training. Training in the investigation of protein-lipid interaction and enzyme mechanisms includes the use of a state-of-the-art computer-driven monolayer apparatus. Analysis of stable isotope incorporation into proteins and lipids in vivo is provided through extensive training in the use of HPLC, GC, and high precision gas isotope ratio mass spectrometry. This practical instruction is coupled with training in the analysis of data using compartmental models to estimate production and transport of proteins and lipids in complex systems. Finally, trainees may receive instruction in the examination of intracellular trafficking of proteins, lipids, and other macromolecules using both upright and inverted confocal fluorescence microscopes coupled to an imaging system with suitable software for analysis of kinetic data. Program faculty participating in providing instruction in these areas are listed below.