Mass spectrometry (MS) based protein footprinting is a new addition to structural biology tool box. The sensitivity and speed of MS provide a bridge between low resolution methods, like circular dichroism, and high resolution methods, like NMR and X-ray crystallography. Footprinting modifies proteins in biologically relevant environments. The modified peptides can be qualitatively and quantitatively analyzed by MS to generate maps of the solvent accessibility of target proteins. This view of the protein structure at the peptide level is sufficient to determine protein conformational changes introduced by the functional switch of proteins and can be investigated without limitations of purity and size. Two approaches are deuterium (hydrogen deuterium exchange, or HDX) and hydroxyl radical labeling (fast photochemical oxidation of proteins, or FPOP). Both of them can label more than half of the amino acids in biologically relevant environments. FPOP can monitor very fast events, down to a microsecond. Phenomena such as fast protein folding, conformational changes, and aggregation can now be probed. In addition, other radicals that react with specific residues will be developed. Combining HDX with MS allows much deeper investigation. Titration-based HDX will be exploited to provide protein-ligand binding affinities. Pulsed HDX will be developed to look at challenging protein aggregates such as amyloid beta (implicated in Alzheimer's Disease). Both technologies will be pushed to determine if they can provide information at the amino acid residue level.

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 #
5P41GM103422-42
Application #
9618639
Study Section
Special Emphasis Panel (ZRG1)
Project Start
Project End
2020-12-31
Budget Start
2019-01-01
Budget End
2019-12-31
Support Year
42
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Washington University
Department
Type
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
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
Xu, Wei; Mukherjee, Sumit; Ning, Yu et al. (2018) Cyclopropane fatty acid synthesis affects cell shape and acid resistance in Leishmania mexicana. Int J Parasitol 48:245-256
Luo, Zonghua; Rosenberg, Adam J; Liu, Hui et al. (2018) Syntheses and in vitro evaluation of new S1PR1 compounds and initial evaluation of a lead F-18 radiotracer in rodents. Eur J Med Chem 150:796-808
Rajagopal, Rithwick; Zhang, Sheng; Wei, Xiaochao et al. (2018) Retinal de novo lipogenesis coordinates neurotrophic signaling to maintain vision. JCI Insight 3:
van Vliet, Stephan; Smith, Gordon I; Porter, Lane et al. (2018) The muscle anabolic effect of protein ingestion during a hyperinsulinaemic euglycaemic clamp in middle-aged women is not caused by leucine alone. J Physiol 596:4681-4692
Keul, Nicholas D; Oruganty, Krishnadev; Schaper Bergman, Elizabeth T et al. (2018) The entropic force generated by intrinsically disordered segments tunes protein function. Nature 563:584-588
Goldner, Nicholas K; Bulow, Christopher; Cho, Kevin et al. (2018) Mechanism of High-Level Daptomycin Resistance in Corynebacterium striatum. mSphere 3:
Zhang, Bojie; Cheng, Ming; Rempel, Don et al. (2018) Implementing fast photochemical oxidation of proteins (FPOP) as a footprinting approach to solve diverse problems in structural biology. Methods 144:94-103
Su, Zhaoming; Wu, Chao; Shi, Liuqing et al. (2018) Electron Cryo-microscopy Structure of Ebola Virus Nucleoprotein Reveals a Mechanism for Nucleocapsid-like Assembly. Cell 172:966-978.e12

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