Abstract
ObjectiveTo characterize the mammalian Fic protein HYPE in terms of its subcellular localization, tissue specific expression, molecular function and potential substratesMethodsClick chemistry, qRT‐PCR, stable cell lines, protein expression and purificationResultsUsing a variety of tools developed in our lab, we have partially characterized the subcellular localization, glycosylation state and tissue specific expression of the mammalian Fic protein termed HYPE. Several forms of the human and mouse HYPE protein have been successfully expressed and purified for use in a variety of assays to determine its potential substrates, and stable cell lines containing HYPE shRNA knockdown and HYPE mutants are being generated in a variety of cell lines to attempt to determine its cellular function and substrates.ConclusionThe significance of the Fic domain activity of mammalian HYPE is not currently known, but our current knowledge, including its subcellular localization and tissues of relevance, combined with our ongoing research with tools like click chemistry, recombinant HYPE protein and stable mammalian cells lines should allow us to elucidate the substrates and functions of this enigmatic protein.
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