Identification of a Genetic Marker of Diabetes Found in the Plasma of Non-Insulin Dependent Diabetic KKAy Mice
Ho, Jeffrey C.
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The purpose of this project was to characterize the KKAy 9kDa protein by obtaining the amino acid content and the amino acid sequence. The sequence will be compared with other known sequences in the protein database to elucidate whether the sequence of the protein has already been described or whether it is a novel protein. Minimal sequence for this type of search is ten amino acid residues. Therefore, only partial sequence of the 9kDa protein is needed (84 residues are predicted using an average molecular mass of 107 Da per amino acid). Since the protein is N-terminally blocked, the protein is to be cleaved either chemically or enzymatically and the resulting polypeptide fragments sequenced using Edman chemistry. Manual gas phase sequencing will be attempted if nanomolar levels of peptide are isolated and automated sequencing will be employed if the fragment yields are on the picomolar level. To gain a more complete understanding of this protein it is useful to run an amino acid analysis to find out the relative amounts of amino acids in the 9kDa protein. Positive characterization of the KKAY 9kDa protein may help indicate if the protein is a precursor or a product of diabetes. It is hypothesized that the 9kDa protein is a genetic marker of diabetes because of its elevated expression level in KKAY mice. However, the protein may not be a 6 causative factor of diabetes but, instead, a secondary diabetic complication. If the protein is found to be a part of the pathogenesis of the disease (whether it be a genetic marker, a precursor, or a product) it may be a useful tool in studying the development and treatment of Type II diabetes.
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