The field of mass spectrometry has developed dramatically in the past few years. Major advances have been made in capillary gas chromatographic-accurate mass mass spectrometry; new ionization techniques such as fast atom bombardment; extended mass range (Greater 1000 daltons) instruments; tandem mass spectrometry (MS/MS); and computer data acquisition and processing. It is now possible to attempt mass spectrometric analysis of important large biological molecules, such as peptide and carbohydrate oligomers.
The aim of this proposal is to apply and further develop new techniques in mass spectrometry to solve specific problems in the chemical analysis of peptide and carbohydrate oligomers. Unmodified and modified peptide oligomers will be obtained from: cytolytic peptides and proteins (melittin; Delta-hemolysin; human eosinophil major basic protein); a calcium dependent bioluminescent protein (aequorin); calcium binding proteins; opiate-binding peptides; cholecystokinin peptides; vitamin K-dependent protein factors; and the cyclic undecapeptide drug cyclosporine. Oligosaccharides analyzed will be of the high mannose type isolated from the urine of patients with mannosidosis; trehalose-containing lipooligosaccharide mycobacterial antigens; and heparin fragments generated by heparinase. Significance: First, significant contributions will be made to the structural and sequence analysis of a number of important biological peptide and saccharide oligomers. Second, the unique applications of new techniques in mass spectrometry to the solutions of these specific problems will be demonstrated. A significant contribution will therefore be made to further the development of new mass spectrometric techniques for the analysis of significant biological oligomers.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032928-03
Application #
3282173
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1984-12-01
Project End
1987-11-30
Budget Start
1986-12-01
Budget End
1987-11-30
Support Year
3
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Mayo Clinic, Rochester
Department
Type
DUNS #
City
Rochester
State
MN
Country
United States
Zip Code
55905
Jardine, I; Scanlan, G; McNeil, M et al. (1989) Plasma desorption mass spectrometric analysis of mycobacterial glycolipids. Anal Chem 61:416-22
Kodaira, H; Lisek, C A; Jardine, I et al. (1989) Identification of the convulsant opiate thebaine in mammalian brain. Proc Natl Acad Sci U S A 86:716-9
Lisek, C A; Bailey, J E; Benson, L M et al. (1989) Quantitation of endogenous substance P by on-line microcolumn liquid chromatography/continuous-flow fast-atom bombardment mass spectrometry. Rapid Commun Mass Spectrom 3:43-6
Tsarbopoulos, A; Gross, M; Kumar, R et al. (1989) Rapid identification of calbindin-D28k cyanogen bromide peptide fragments by plasma desorption mass spectrometry. Biomed Environ Mass Spectrom 18:387-93
Hunter, S W; Barr, V L; McNeil, M et al. (1988) Trehalose-containing lipooligosaccharide antigens of Mycobacterium sp.: presence of a mono-O-methyltri-O-acyltrehalose ""core"". Biochemistry 27:1549-56
Tsarbopoulos, A; Becker, G W; Occolowitz, J L et al. (1988) Peptide and protein mapping by 252Cf-plasma desorption mass spectrometry. Anal Biochem 171:113-23
Jardine, I; Scanlan, G F; Tsarbopoulos, A et al. (1988) Plasma desorption mass spectrometry of peptides adsorbed on nitrocellulose from a glutathione matrix. Anal Chem 60:1086-8
Bozic, C M; McNeil, M; Chatterjee, D et al. (1988) Further novel amido sugars within the glycopeptidolipid antigens of Mycobacterium avium. J Biol Chem 263:14984-91
Donnerer, J; Cardinale, G; Coffey, J et al. (1987) Chemical characterization and regulation of endogenous morphine and codeine in the rat. J Pharmacol Exp Ther 242:583-7
McNeil, M; Tsang, A Y; McClatchy, J K et al. (1987) Definition of the surface antigens of Mycobacterium malmoense and use in studying the etiology of a form of mycobacteriosis. J Bacteriol 169:3312-20

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