Abstract

This proposal describes a program for investigating the conformations of gas-phase biomolecules and complexes. Preliminary data show that when ESI is used to produce gas-phase biomolecules, Coulombic forces that result from the multiple charges appear to be a dominant factor in establishing structure in the gas phase. In order to explore the role of noncovalent interactions in establishing structure it is proposed to develop a hybrid drift tube/time-of-flight mass spectrometer for studying high-mass/low-charge biomolecules and complexes. Questions regarding the number of different gas-phase conformations, their shapes and the connectivity between them will be addressed. Although these methods cannot give atomic detail, it is possible to extract overall shape information. The methods proposed should be fast and sensitive and make it possible to study trace amount of biological materials. The ability to study biomolecular structure in vacuum allows molecular issue that are fundamental to biochemistry, such as folding recognition, biomolecular association, and the influence of solvent on structure to be explored experimentally. The raw data will provide benchmarks for theorists who aim to calculate protein structures and dynamics in vacuo and in solution.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM055647-02
Application #
2701821
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1997-05-01
Project End
2000-04-30
Budget Start
1998-05-01
Budget End
1999-04-30
Support Year
2
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
Indiana University Bloomington
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
006046700
City
Bloomington
State
IN
Country
United States
Zip Code
47401

  Publications

Counterman, A E; Clemmer, D E (2001) Large anhydrous polyalanine ions: evidence for extended helices and onset of a more compact state. J Am Chem Soc 123:1490-8
Lee, Y J; Hoaglund-Hyzer, C S; Taraszka, J A et al. (2001) Collision-induced dissociation of mobility-separated ions using an orifice-skimmer cone at the back of a drift tube. Anal Chem 73:3549-55
Srebalus, C A; Li, J; Marshall, W S et al. (2000) Determining synthetic failures in combinatorial libraries by hybrid gas-phase separation methods. J Am Soc Mass Spectrom 11:352-5
Srebalus, C A; Li, J; Marshall, W S et al. (1999) Gas-phase separations of electrosprayed peptide libraries. Anal Chem 71:3918-27
Valentine, S J; Counterman, A E; Clemmer, D E (1999) A database of 660 peptide ion cross sections: use of intrinsic size parameters for bona fide predictions of cross sections. J Am Soc Mass Spectrom 10:1188-211
Henderson, S C; Valentine, S J; Counterman, A E et al. (1999) ESI/ion trap/ion mobility/time-of-flight mass spectrometry for rapid and sensitive analysis of biomolecular mixtures. Anal Chem 71:291-301
Hoaglund, C S; Valentine, S J; Sporleder, C R et al. (1998) Three-dimensional ion mobility/TOFMS analysis of electrosprayed biomolecules. Anal Chem 70:2236-42
Valentine, S J; Counterman, A E; Hoaglund, C S et al. (1998) Gas-phase separations of protease digests. J Am Soc Mass Spectrom 9:1213-6
Counterman, A E; Valentine, S J; Srebalus, C A et al. (1998) High-order structure and dissociation of gaseous peptide aggregates that are hidden in mass spectra. J Am Soc Mass Spectrom 9:743-59