The broad long term goal of this research is to develop a new mass spectrometric method for the structural elucidation of oligosaccharides. Oligosaccharides (OS) play a central role in mammalian biology in many fundamental processes such as cell-cell recognition, adhesiveness, control of cell division, cellular differentiation and malignant transformation. Hence, in the study of cancer and other related diseases, a general, sensitive, and accurate method of analysis is clearly desired. Mass spectrometry has the potential to offer many of these features. Our immediate goal is to understand the production and unimolecular chemistry of complex gas-phase oligosaccharide ions. The newly developed external source Fourier transform mass spectrometry instrument will be utilized and further developed for the specific analysis of these compounds. This method is unique and allows ions to be produced and trapped for long periods of time. The addition of the external source has allowed the use of fast atom bombardment to generate gas-phase bio-organic ions. Preliminary experiments have been performed showing major differences between this technique and other mass spectrometric methods. We find that matrix interference in the spectra is minimal add substantial fragmentation is obtained. This is due to the relatively longer detection time scale of the instrument. During the detection period, slow metastable decay occurs and allows oligosaccharide ions to undergo fragmentation. Matrix cluster ions also undergo metastable decomposition. However, these ions fragment more quickly due to the loose nature of the bonding. The result is a spectrum of oligosaccharide ions in the near absence of matrix interference. Studies are proposed to gain a better understanding of the unimolecular decay. In addition, conditions will be obtained to optimize structural information from FAB/FTMS alone (i.e. without further tandem MS determinations). This would make powerful the coupling of liquid chromatography techniques such as flow FAB (and electrospray) to this mass spectral method to analyze complex oligosaccharide mixtures. Direct application to glycolipids and high-mannose compounds will be performed through a collaboration with a natural products chemist and an oligosaccharide biochemists.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29GM049077-02
Application #
2186613
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1993-05-01
Project End
1998-04-30
Budget Start
1994-05-01
Budget End
1995-04-30
Support Year
2
Fiscal Year
1994
Total Cost
Indirect Cost
Name
University of California Davis
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
094878337
City
Davis
State
CA
Country
United States
Zip Code
95618
Park, Dayoung; Brune, Kristin A; Mitra, Anupam et al. (2015) Characteristic Changes in Cell Surface Glycosylation Accompany Intestinal Epithelial Cell (IEC) Differentiation: High Mannose Structures Dominate the Cell Surface Glycome of Undifferentiated Enterocytes. Mol Cell Proteomics 14:2910-21
De Leoz, Maria Lorna A; Kalanetra, Karen M; Bokulich, Nicholas A et al. (2015) Human milk glycomics and gut microbial genomics in infant feces show a correlation between human milk oligosaccharides and gut microbiota: a proof-of-concept study. J Proteome Res 14:491-502
Guerrero, Andres; Dallas, David C; Contreras, Stephanie et al. (2014) Mechanistic peptidomics: factors that dictate specificity in the formation of endogenous peptides in human milk. Mol Cell Proteomics 13:3343-51
Wu, Shuai; Salcedo, Juli; Tang, Ning et al. (2012) Employment of tandem mass spectrometry for the accurate and specific identification of oligosaccharide structures. Anal Chem 84:7456-62
Nwosu, Charles C; Aldredge, Danielle L; Lee, Hyeyoung et al. (2012) Comparison of the human and bovine milk N-glycome via high-performance microfluidic chip liquid chromatography and tandem mass spectrometry. J Proteome Res 11:2912-24
Lee, Hyeyoung; An, Hyun Joo; Lerno Jr, Larry A et al. (2011) Rapid Profiling of Bovine and Human Milk Gangliosides by Matrix-Assisted Laser Desorption/Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry. Int J Mass Spectrom 305:138-150
Lerno Jr, Larry A; German, J Bruce; Lebrilla, Carlito B (2010) Method for the identification of lipid classes based on referenced Kendrick mass analysis. Anal Chem 82:4236-45
Kronewitter, Scott R; de Leoz, Maria Lorna A; Peacock, Kyle S et al. (2010) Human serum processing and analysis methods for rapid and reproducible N-glycan mass profiling. J Proteome Res 9:4952-9
Dodds, Eric D; Tassone, Flora; Hagerman, Paul J et al. (2009) Polymerase chain reaction, nuclease digestion, and mass spectrometry based assay for the trinucleotide repeat status of the fragile X mental retardation 1 gene. Anal Chem 81:5533-40
Seipert, Richard R; Dodds, Eric D; Lebrilla, Carlito B (2009) Exploiting differential dissociation chemistries of O-linked glycopeptide ions for the localization of mucin-type protein glycosylation. J Proteome Res 8:493-501

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