Semester

Spring

Date of Graduation

2004

Document Type

Thesis

Degree Type

MS

College

Eberly College of Arts and Sciences

Department

Chemistry

Committee Chair

Aaron Timperman.

Abstract

Protein structure and function are often regulated by covalent modifications to the protein. One of the most important of these modifications is phosphorylation, which can control many cellular processes. The sites of phosphorylation must be determined to fully understand the protein's function and regulatory events, which can be challenging because of the low stoichiometry of the phosphorylated regulatory proteins.;Several strategies for enrichment of phosphopeptides prior to analysis have been developed, however, Immobilized Metal Affinity Chromatography (IMAC) is the most widely used method. Online IMAC-ESI-MS has been developed with ammonium hydroxide as the elutor. Gradient elution from the IMAC column demonstrates that the phosphopeptides were found to desorb from the IMAC slowly, leading to broad peak widths. By increasing the temperature, the elution from the IMAC column was more rapid, which made the peak widths more narrow.

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