Mass spectrometry has become the enabling tool for identification of both lipids and proteins and for determination of their structures and properties. The changes in lipids with disease have become increasingly recognized, suggesting they may be biomarkers. One goal of the Washington University Mass Spectrometry Research Resource is to build a foundation for understanding lipid fragmentation, facilitating both de novo and computer-assisted identification of increasingly complex lipids found in decreasing amounts in bacteria and parasites. This work will continue because we recognize the growing challenge to identify more complex lipids and to uncover their role in infection and disease. In protein science and proteomics, related challenges exist to understand, with greater speed and sensitivity, protein properties and interactions. As a second goal, various chemical footprinting procedures that take advantage ofthe separation and analysis capabilities of modern chromatography and mass spectrometry will be explored as sources of information on protein structure, folding, and interfaces with metal ions, DNA, and other proteins. The strategy is to build a """"""""tool box"""""""" of footprinting reagents that overtly modify proteins to map those regions that are accessible to reaction, thereby revealing interfaces and quantifying affinities. This approach takes full advantage of the power of modern """"""""bottom-up"""""""" proteomics and is designed to be exportable to any proteomics laboratory. Nevertheless, bottom-up analytical proteomics fails when confronted with the need to identify the complex arrays of protein modifications that often form in post-translational modification or are introduced by footprinting. Addressing this problem by improving top-down proteomics is the third goal of the WU Resource. We propose to develop new instrumentation, methods, and associated data processing and interpretation strategies. They will be implemented along with a new high-field Fourier transform ion cyclotron resonance mass spectrometer to aid biomedical researchers with problems for which protein modification is an issue. Its success will serve to demonstrate a paradigm for top-down sequencing that can be employed by other researchers who search for the fine details of protein structure and function.

Public Health Relevance

Lipids and protein are important components in all living systems. Identifying them, characterizing their properties and interactions with other substances, and determining how they can be altered is important for understanding human health and disease. Mass spectrometry is meeting these challenges and moving forward with the innovative instrumentation and methods proposed here.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Biotechnology Resource Grants (P41)
Project #
Application #
Study Section
Special Emphasis Panel (ZRG1-BCMB-K (40))
Program Officer
Sheeley, Douglas
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Washington University
Internal Medicine/Medicine
Schools of Medicine
Saint Louis
United States
Zip Code
Shen, Guomin; Li, Shuang; Cui, Weidong et al. (2018) Membrane Protein Structure in Live Cells: Methodology for Studying Drug Interaction by Mass Spectrometry-Based Footprinting. Biochemistry 57:286-294
Mukherjee, Sumit; Xu, Wei; Hsu, Fong-Fu et al. (2018) Sterol methyltransferase is required for optimal mitochondrial function and virulence in Leishmania major. Mol Microbiol :
Sato, Chihiro; Barthélemy, Nicolas R; Mawuenyega, Kwasi G et al. (2018) Tau Kinetics in Neurons and the Human Central Nervous System. Neuron 97:1284-1298.e7
Rocha, Agostinho G; Franco, Antonietta; Krezel, Andrzej M et al. (2018) MFN2 agonists reverse mitochondrial defects in preclinical models of Charcot-Marie-Tooth disease type 2A. Science 360:336-341
Frankfater, Cheryl; Jiang, Xuntian; Hsu, Fong-Fu (2018) Characterization of Long-Chain Fatty Acid as N-(4-Aminomethylphenyl) Pyridinium Derivative by MALDI LIFT-TOF/TOF Mass Spectrometry. J Am Soc Mass Spectrom 29:1688-1699
Henson, William R; Hsu, Fong-Fu; Dantas, Gautam et al. (2018) Lipid metabolism of phenol-tolerant Rhodococcus opacus strains for lignin bioconversion. Biotechnol Biofuels 11:339
Lucey, Brendan P; Hicks, Terry J; McLeland, Jennifer S et al. (2018) Effect of sleep on overnight cerebrospinal fluid amyloid ? kinetics. Ann Neurol 83:197-204
Johnson, Britney; McConnell, Patrick; Kozlov, Alex G et al. (2018) Allosteric Coupling of CARMIL and V-1 Binding to Capping Protein Revealed by Hydrogen-Deuterium Exchange. Cell Rep 23:2795-2804
Shu, Longfei; Zhang, Bojie; Queller, David C et al. (2018) Burkholderia bacteria use chemotaxis to find social amoeba Dictyostelium discoideum hosts. ISME J 12:1977-1993
Zayed, Mohamed A; Hsu, Fong-Fu; Patterson, Bruce W et al. (2018) Diabetes adversely affects phospholipid profiles in human carotid artery endarterectomy plaques. J Lipid Res 59:730-738

Showing the most recent 10 out of 323 publications