Selection and analysis of phosphopeptides in proteomics
Abstract
Protein phosphorylation is a very important post-translational modification that has been implicated in many processes such as signal transduction, cell growth, disease, along with several others. Accordingly, much effort has been put forth to study the sites and extent of phosphorylation on proteins and peptides. Ga(III) IMAC combined with digestion using glu-C instead of the more traditional trypsin was examined for its ability to reduce the nonspecific binding typically associated with IMAC selection. β-elimination/Michael addition was used in conjunction with diagonal chromatography and strong cation exchange chromatography as alternate means for selection and identification of phosphopeptides. Upon determination that Ga(III) IMAC selection with glu-C digestion was the superior method, it was used to study the changes in phosphorylation of phosphatase inhibitor liver toxin, Microcystin-LR, treated mouse livers. Ga(III) IMAC selection and alkaline phosphatase treatment were used in the study of relative amounts of different phosphoforms of a protein implicated in cancer.
Degree
Ph.D.
Advisors
Regnier, Purdue University.
Subject Area
Analytical chemistry|Biochemistry
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