Peptide Fragmentation and Amino Acid Quantification by Mass Spectrometry

Research presented in this dissertation falls into two parts: fragmentation mechanisms of peptide and fragmentation mechanism of amino acid derivatives. The study of peptide fragmentation may help to improve protein identification by incorporating the rules governing this process into search algorit...

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Bibliographic Details
Main Author: Zhang, Qingfen
Other Authors: Wysocki, Vicki H.
Language:en
Published: The University of Arizona. 2006
Subjects:
Online Access:http://hdl.handle.net/10150/195288
Description
Summary:Research presented in this dissertation falls into two parts: fragmentation mechanisms of peptide and fragmentation mechanism of amino acid derivatives. The study of peptide fragmentation may help to improve protein identification by incorporating the rules governing this process into search algorithms. This study elucidates the chemical 'rules' governing peptide dissociation. It is believed that these 'rules' can be incorporated into searching algorithms to achieve better protein identification. The present study focuses on the effects of different amino acids on fragmentation. Amino acids with a wide range of different chemical and physical properties are investigated, including amino acids with hydrophilic side chains, amino acids with aliphatic side chains and amino acids without side chains. It can be concluded from the present studies that the different amino acid properties have great influence on the peptide fragmentation and spectrum appearance.The study of fragmentation mechanisms of amino acid derivatives is another focus of this dissertation. Based on the fragmentation mechanism study, a quantification method was developed. The method can distinguish glutamine with 15N-label at N-terminal amine vs the side chain even if they have same molecular weight. Ammonia metabolism was successfully monitored by feeding mosquitoes with isotope-labeled compounds and subsequently measuring the amount of the labeled amino acids. This method demonstrates the power of mass spectrometry in metabolism studies.