Tandem mass spectrometry (MS/MS, MSn) employing a Fourier-transform mass spectrometer with electrospray ionization will be applied to large biomolecules. Key objectives will be to obtain MSn data with high mass accuracy (errors <1 ppm) and high resolution (unit resolution or higher) for molecules of molecular weight >10 kDa, possibly up to 150-200 kDa. Fourier and Hadamard transform capabilities will allow all such MS and MS/MS, respectively, data to be collected simultaneously over a broad mass range (m/z >200) on femtomole or subfemtomole sample quantities. Ion cell improvements contributing to these higher capabilities will include ion excitation methods, rf excitation field uniformly, and electrostatic field homogeneity. Improved computer hardware and software will more efficiently utilize this unusually high volume of data. Specific ion-molecule reactions will be applied to the MS/MS characterization of these large multiply-charged ions. Applications studied will include surveying unknown biological system constituents, detailed structural characterization of biomolecules (sequences, adducts, S-S bridge linkages), routine assays of recombinant proteins, and quantitation of enzyme-substrate and receptor- ligand affinities. Fundamental research on unimolecular ion dissociations will be extended to large multiply-charged positive and negative ions, including the kinetic redistribution of excitation energy. MS information will be sought indicative of three-dimensional structures of gaseous biomolecule ions and protein folding.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Method to Extend Research in Time (MERIT) Award (R37)
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Special Emphasis Panel (NSS)
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Edmonds, Charles G
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Cornell University
Schools of Arts and Sciences
United States
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Skinner, Owen S; Breuker, Kathrin; McLafferty, Fred W (2013) Charge site mass spectra: conformation-sensitive components of the electron capture dissociation spectrum of a protein. J Am Soc Mass Spectrom 24:807-10
Begley, Tadhg P; Ealick, Steven E; McLafferty, Fred W (2012) Thiamin biosynthesis: still yielding fascinating biological chemistry. Biochem Soc Trans 40:555-60
Breuker, Kathrin; Skinner, Owen S; McLafferty, Fred W (2012) Femtosecond laser vaporization that preserves protein-folded structure: an unproven idea. Proc Natl Acad Sci U S A 109:E206; author reply E207
Skinner, Owen S; McLafferty, Fred W; Breuker, Kathrin (2012) How ubiquitin unfolds after transfer into the gas phase. J Am Soc Mass Spectrom 23:1011-4
Kong, Xianglei; Lin, Cheng; Infusini, Giuseppe et al. (2009) Numerous isomers of serine octamer ions characterized by infrared photodissociation spectroscopy. Chemphyschem 10:2603-6
Han, Xuemei; Smith, Norah L; Sil, Dwaipayan et al. (2009) IgE receptor-mediated alteration of membrane-cytoskeleton interactions revealed by mass spectrometric analysis of detergent-resistant membranes. Biochemistry 48:6540-50
Breuker, Kathrin; McLafferty, Fred W (2008) Stepwise evolution of protein native structure with electrospray into the gas phase, 10(-12) to 10(2) s. Proc Natl Acad Sci U S A 105:18145-52
McLafferty, Fred W (2008) Mass spectrometry across the sciences. Proc Natl Acad Sci U S A 105:18088-9
Narayan, Mahesh; Welker, Ervin; Zhai, Huili et al. (2008) Detecting native folds in mixtures of proteins that contain disulfide bonds. Nat Biotechnol 26:427-9
McLafferty, Fred W; Breuker, Kathrin; Jin, Mi et al. (2007) Top-down MS, a powerful complement to the high capabilities of proteolysis proteomics. FEBS J 274:6256-68

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