An instrument will be developed for the mass spectral analysis of biomolecules which combines the best features of high flux particle bombardment (large secondary ion currents, the use of a liquid matrix and solution chemistry) and time-of-flight analysis (low cost and high ion transmission). High speed integrated transient recording will be used rather than the pulse counting techniques currently in use with plasma desorption and low flux SIMS time-of-flight configurations. The design will permit a direct comparison of the secondary ion spectra (molecular ions vs. fragmentation) under low and high primary ion flux conditions, with particular attention to the question of metastable decompositions from both methods. Time delayed ion extraction techniques, developed previously for laser desorption, will be used here to enhance high mass, low energy ion formation and focussing. The instrument will be applied to problems in biochemistry where rapid routine analysis can be undertaken under low (mass) resolution conditions, such as qualitative identification of molecular ions of peptides in hydrolysis mixtures, components of the lipid-A complex and mixtures of glycopeptides, or sequence changes in interspecies comparisons of epidermal growth factors. This proposal is identical with a proposal (GM 33967-01) submitted previously, except that: (1) results are reported on a feasibility study which demonstrates stable secondary ion currents on a time-of-flight using high flux primary beams (Section C), and (2) instrumental details are provided, based upon the use of a pulsed inert gas ion gun (Section D).

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM033967-02
Application #
3284229
Study Section
(BET)
Project Start
1985-09-01
Project End
1988-08-31
Budget Start
1986-09-01
Budget End
1987-08-31
Support Year
2
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Johns Hopkins University
Department
Type
Schools of Medicine
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Smit, John; Kaltashov, Igor A; Kaltoshov, Igor A et al. (2008) Structure of a novel lipid A obtained from the lipopolysaccharide of Caulobacter crescentus. Innate Immun 14:25-37
Cotter, R J; Fancher, C; Cornish, T J (1999) Miniaturized time-of-flight mass spectrometer for peptide and oligonucleotide analysis. J Mass Spectrom 34:1368-72
Dotson, G D; Kaltashov, I A; Cotter, R J et al. (1998) Expression cloning of a Pseudomonas gene encoding a hydroxydecanoyl-acyl carrier protein-dependent UDP-GlcNAc acyltransferase. J Bacteriol 180:330-7
Odegaard, T J; Kaltashov, I A; Cotter, R J et al. (1997) Shortened hydroxyacyl chains on lipid A of Escherichia coli cells expressing a foreign UDP-N-acetylglucosamine O-acyltransferase. J Biol Chem 272:19688-96
Cornish, T J; Cotter, R J (1997) High-order kinetic energy focusing in an end cap reflectron time-of-flight mass spectrometer. Anal Chem 69:4615-8
White, K A; Kaltashov, I A; Cotter, R J et al. (1997) A mono-functional 3-deoxy-D-manno-octulosonic acid (Kdo) transferase and a Kdo kinase in extracts of Haemophilus influenzae. J Biol Chem 272:16555-63
Kaltashov, I A; Doroshenko, V; Cotter, R J et al. (1997) Confirmation of the structure of lipid A derived from the lipopolysaccharide of Rhodobacter sphaeroides by a combination of MALDI, LSIMS, and tandem mass spectrometry. Anal Chem 69:2317-22
Kaltashov, I A; Doroshenko, V M; Cotter, R J (1997) Gas phase hydrogen/deuterium exchange reactions of peptide ions in a quadrupole ion trap mass spectrometer. Proteins 28:53-8
Cordero, M M; Cornish, T J; Cotter, R J et al. (1995) Sequencing peptides without scanning the reflectron: post-source decay with a curved-field reflectron time-of-flight mass spectrometer. Rapid Commun Mass Spectrom 9:1356-61
Fabris, D; Vestling, M M; Cordero, M M et al. (1995) Sequencing electroblotted proteins by tandem mass spectrometry. Rapid Commun Mass Spectrom 9:1051-5

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