Application of Mass Spectrometry to Structural Biology

Tomer, K B.

Abstract

Recent instrumental developments in mass spectrometry, such as electrospray ionization (ESI) and matrix assisted laser desorption time-of- flight mass spectrometry (MALDI TOF), have enabled the mass spectrometrist to investigate biological species with significantly higher average molecular weight than previously possible. The combination of these techniques with chemical processing of large biological species such as proteins now enables the use of mass spectrometry to determine a wide range of problems within the realm of structural biology. Within the past year, we have begun to address several issues in structural biochemistry. The goal of one project, in collaboration with Dr J. Hoeg, MDB/NHLBI/NIH, and W. Weinmann/ Prof. M. Przybylski, Univ. Konstanz, is to determine the sites of palmitoylation analon of phosphorylation of apolipoprotein A-1 secreted by the human hepatoma cell line Hep G2. A second issue is the isolation/structure determination of proteins that have been conjugated with xenobiotics through exposure. The first model to be investigated in this system is the hemoglobin- isoprene adduct isolated from the blood of mice exposed to isoprene. The third issue being addressed is the application of MS techniques to the investigation of enzyme-substrate/receptor-substrate interactions.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of Environmental Health Sciences (NIEHS)
• Type: Intramural Research (Z01)
• Project #: 1Z01ES050127-01
• Application #: 3777545
• Support Year: 1
• Fiscal Year: 1993

Institution

• Name: National Institute of Environmental Health Sciences
• Country: United States

Publications

Schorzman, Allison N; Perera, Lalith; Cutalo-Patterson, Jenny M et al. (2011) Modeling of the DNA-binding site of yeast Pms1 by mass spectrometry. DNA Repair (Amst) 10:454-65

Dhungana, Suraj; Fessler, Michael B; Tomer, Kenneth B (2009) Epitope mapping by differential chemical modification of antigens. Methods Mol Biol 524:119-34

Perdivara, Irina; Deterding, Leesa; Moise, Adrian et al. (2008) Determination of primary structure and microheterogeneity of a beta-amyloid plaque-specific antibody using high-performance LC-tandem mass spectrometry. Anal Bioanal Chem 391:325-36

Iacob, Roxana E; Keck, Zhenyong; Olson, Oakley et al. (2008) Structural elucidation of critical residues involved in binding of human monoclonal antibodies to hepatitis C virus E2 envelope glycoprotein. Biochim Biophys Acta 1784:530-42

Sharp, Joshua S; Tomer, Kenneth B (2007) Analysis of the oxidative damage-induced conformational changes of apo- and holocalmodulin by dose-dependent protein oxidative surface mapping. Biophys J 92:1682-92

Prasad, Rajendra; Liu, Yuan; Deterding, Leesa J et al. (2007) HMGB1 is a cofactor in mammalian base excision repair. Mol Cell 27:829-41

Venkatesh, Sanjay; Tomer, Kenneth B; Sharp, Joshua S (2007) Rapid identification of oxidation-induced conformational changes by kinetic analysis. Rapid Commun Mass Spectrom 21:3927-36

Clark, Alan B; Deterding, Leesa; Tomer, Kenneth B et al. (2007) Multiple functions for the N-terminal region of Msh6. Nucleic Acids Res 35:4114-23

Robinette, David; Neamati, Nouri; Tomer, Kenneth B et al. (2006) Photoaffinity labeling combined with mass spectrometric approaches as a tool for structural proteomics. Expert Rev Proteomics 3:399-408

Sharp, Joshua S; Sullivan, Daniel M; Cavanagh, John et al. (2006) Measurement of multisite oxidation kinetics reveals an active site conformational change in Spo0F as a result of protein oxidation. Biochemistry 45:6260-6