The importance of the structure-function relationship of proteins and protein-complexes motivates the direct measurement of protein structure in the gas phase by mass spectrometry. This method, termed native mass spectrometry or native electrospray ionization, allows the study of intact complexes and the determination of stoichiometry. The native proteins and complexes can also be investigated by a variety of activation methods to determine structural information. These methods include collisionally activated dissociation (CAD), electron capture/transfer dissociation (ECD/ETD), and infrared multi- photon dissociation (IRMPD). Among the systems we will interrogate using these methods are: (1) Lipoprotein complexes, such as high-density lipoproteins (HDLs) whose heterogeneity is of interest. HDL is an important indicator of cardiovascular health. (2) Membrane-embedded proteins play key roles in cell biology and are pharmaceutical targets (e.g., in anesthesia). (3) Proteins with flexible regions are often actively involved in function and interfacing to other proteins. (4) The stoichiometry of very large protein assemblies is among the first information needed to understand system function and it is still very difficult to get for large complexes. (5) Crosslinking studies provide distance constraints and aid in modeling the geometries of large complexes.

Public Health Relevance

-Public Health Relevance. The Washington University Biomedical Mass Spectrometry Resource has a longstanding history as an active and productive citizen in the NIH Biotechnology Research Resources community. We propose to extend our mission by advancing mass spectrometry technology, development, and research, applying these discoveries to answer critical biomedical research questions, and training the next generation of researchers, towards the ultimate goal of improving public health.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Biotechnology Resource Grants (P41)
Project #
5P41GM103422-41
Application #
9412482
Study Section
Special Emphasis Panel (ZRG1)
Project Start
Project End
Budget Start
2018-01-01
Budget End
2018-12-31
Support Year
41
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Washington University
Department
Type
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Yue, Xuyi; Jin, Hongjun; Luo, Zonghua et al. (2017) Chiral resolution of serial potent and selective ?1 ligands and biological evaluation of (-)-[18F]TZ3108 in rodent and the nonhuman primate brain. Bioorg Med Chem 25:1533-1542
Wallen, Jamie R; Zhang, Hao; Weis, Caroline et al. (2017) Hybrid Methods Reveal Multiple Flexibly Linked DNA Polymerases within the Bacteriophage T7 Replisome. Structure 25:157-166
Domingo-Gonzalez, Racquel; Das, Shibali; Griffiths, Kristin L et al. (2017) Interleukin-17 limits hypoxia-inducible factor 1? and development of hypoxic granulomas during tuberculosis. JCI Insight 2:
Frieden, Carl; Wang, Hanliu; Ho, Chris M W (2017) A mechanism for lipid binding to apoE and the role of intrinsically disordered regions coupled to domain-domain interactions. Proc Natl Acad Sci U S A 114:6292-6297
Reeds, Dominic N; Pietka, Terri A; Yarasheski, Kevin E et al. (2017) HIV infection does not prevent the metabolic benefits of diet-induced weight loss in women with obesity. Obesity (Silver Spring) 25:682-688
Wang, Hanliu; Shu, Qin; Frieden, Carl et al. (2017) Deamidation Slows Curli Amyloid-Protein Aggregation. Biochemistry 56:2865-2872
Lu, Yue; Liu, Haijun; Saer, Rafael G et al. (2017) Native Mass Spectrometry Analysis of Oligomerization States of Fluorescence Recovery Protein and Orange Carotenoid Protein: Two Proteins Involved in the Cyanobacterial Photoprotection Cycle. Biochemistry 56:160-166
Zhang, Hao; Harrington, Lucas B; Lu, Yue et al. (2017) Native Mass Spectrometry Characterizes the Photosynthetic Reaction Center Complex from the Purple Bacterium Rhodobacter sphaeroides. J Am Soc Mass Spectrom 28:87-95
Ohlemacher, Shannon I; Giblin, Daryl E; d'Avignon, D André et al. (2017) Enterobacteria secrete an inhibitor of Pseudomonas virulence during clinical bacteriuria. J Clin Invest 127:4018-4030
Niu, Ben; Mackness, Brian C; Rempel, Don L et al. (2017) Incorporation of a Reporter Peptide in FPOP Compensates for Adventitious Scavengers and Permits Time-Dependent Measurements. J Am Soc Mass Spectrom 28:389-392

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