This is an application for a Scientist DevelopmentAward. The long term objectives of this proposal are to establish thepossible role of ALDH5 in acetaldehyde metabolism. This will involvepurification and physical characterization of the ALDH5 protein. Theseobjectives will lead to studies to ascertain the mechanism of liver- specific ALDH5 gene expression. The hypothesis that ALDH5 plays asignificant role in acetaldehyde metabolism stems from preliminaryexperiments which indicate that ALDH5 has a low Km -20 uM for shortchain aldehydes. In addition there is a high degree of homology betweenALDH5 and ALDH2, the major acetaldehyde metabolizing enzyme. Characterization of ALDH5 will involve purification of the recombinantbacterially expressed protein, determination of its molecular weight, determination of various kinetic parameters and substrate preference, determination of subcellular localization, and an examination of thepromoter elements responsible for liver-specific gene expression. Theimmediate objectives of this application will continue work whichinvolves ALDH2 gene expression, which centers around the purificationof a novel liver-specific transcription factor. I have had a long standing interest in alcohol research which began withmy thesis work studying the transcriptional regulation of the humanClass I alcohol dehydrogenase genes. A career development award wouldallow me to develop expertise in protein biochemistry and establish anindependent alcohol-related research program. Studying the structureand function of ALDH5 as well as characterization of a noveltranscription factor involved in ALDH2 gene expression would be alogical direction for me to take in developing into an independentinvestigator.
Acetaldehyde, Aldehyde Dehydrogenase, Enzyme Substrate, Gene Expression, Genetic Promoter Element, Laboratory Rabbit, Laboratory Rat, Liver Cell, Metabolism, Mitochondria, Molecular Weight, Protein Purification, Recombinant Protein, Transcription Factor
