Candidate: The candidate is presently a Research Scientist and has already been approved for promotion to Assistant Research Professor by Dr. A. Wasserman, Chairman of the Dept. of Medicine at GWU. He has been working in the sponsor's laboratory for the past 6 years (8 articles, 2 reviews). Immediate and long-term career goals: apply for Associate Professorship and Full Professorship at the Department of Medicine, George Washington University. Expert consultant in the fields of protein and lipid biochemistry. Research career development plan: work on the proposed grant project and simultaneously develop, explore and train in related topics in alcoholism research to establish himself as an independent investigator. Apply for independent investigator awards for funding his projects involving the G-proteins (trafficking) study and others in which he was involved, such as the effects of alcohol on brain gangliosides and behavior, enzymology of beta-carotene metabolism. Environment: The sponsor's laboratory (2,250 sq.ft lab space, 320 sq.ft animal facility, 1000 sq.ft common instrument room) is located in the Research Building of the VA Medical Center in Washington, DC. It has an ideal environment for both basic and clinical research. The sponsor has two NIAAA funded projects and one NCI funded project. Research: Ethanol disrupts liver intracellular protein trafficking resulting in the accumulation of nascent proteins, as well as lipids that have to be assembled and transported as lipoproteins. However, the pathogenic mechanisms of action of ethanol are not well understood. Thus, to understand the underlying causes for alcohol mediated accumulation of proteins and lipids, we plan to elucidate the mechanism of action of ethanol on their post-translational modification and trafficking with special emphasis on small GTP binding proteins (rabs) that are crucial in the vesicular transport process. Current research and our preliminary data raise the following questions: 1). Does ethanol affect intracellular distribution of newly synthesized export proteins? 2). What are the sites at which ethanol alters intracellular protein trafficking? 3). Does ethanol affect specific rab proteins in protein trafficking? In the present study, the Candidate will attempt to answer these questions, using transferrin (N-glycosylated), apoE (O-glycosylated), VLDL (triglycerides carrier) and albumin (non-glycosylated) as model liver proteins. Therefore, the Candidate will study the effects of chronic ethanol treatment on model liver proteins at the following levels: intracellular distribution (aims 1,4), intracellular trafficking (aims 2,5) and the disposition of prenylated rabs attached to newly synthesized model liver proteins in various subcellular organelles (aims 3,6). All studies will be done as a function of concentration and time of ethanol exposure in rat liver in vivo (aims 1-3) or human liver in vitro in wild type HepG2 cells, HAD cells and CYP2E1 cells (aims 4-6).
