Several problems in pharmacology will be addressed, in which significant progress can be made by determining the primary sequence of proteins or altered proteins. The speed and accuracy of mass spectrometry will be brought to these problems, in conjunction with chemical and enzymatic techniques. New mass spectrometry techniques will be developed in support of protein structure studies. We will continue studies directed at understanding the multiple mechanisms for the drug resistance often acquired during chemotherapy, by characterizing the covalent sequestration proposed for both induced metallothionein and induced glutathione-Stransferase. Alkylation of purified proteins will be characterized for three therapeutic mustards, which have a range of chemical reactivities. In addition, sites of covalent bonding by activated glucuronides on human serum albumin and on cytosolic glutathione-S-transferases will be defined. Identification of modified residues may provide new information about binding sites for these widely occurring metabolites, and provide a molecular mechanism for the idiopathic immunogenicity associated with nonsteroidal anti-inflammatory and other drugs. Polyethylene glycol-derivatized adenosine deaminase (PEG-ADA) is an orphan drug used in treatment of severe immunosuppression syndrome. Analytical methods will be developed to characterize this very complex mixture, to understand the structural determinants of its plasma stability and immunocompatibility, and to enable metabolism studies. These will be applicable to other PEGated proteins under development for therapeutic uses. In combination these studies will provide insight as to whether xenobiotic alkylation of serum and cytosolic proteins is random or ordered, and how structural features in the drug and protein influence selectivity. Mass spectrometry techniques under development in support of these studies of pharmacologically relevant proteins include reaction induced dissociation in our high performance tandem mass spectrometer, and electrospray ionization with collisional activation or reaction induced dissociation.

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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Metallobiochemistry Study Section (BMT)
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University of Maryland Baltimore
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Rose, Rebecca L; Choksawangkarn, Waeowalee; Fenselau, Catherine (2018) Application of Higher Density Iron Oxide Nanoparticle Pellicles to Enrich the Plasma Membrane and Its Proteome from Cells in Suspension. Methods Mol Biol 1722:79-90
Chen, Dapeng; Gomes, Fabio; Abeykoon, Dulith et al. (2018) Top-Down Analysis of Branched Proteins Using Mass Spectrometry. Anal Chem 90:4032-4038
Geis-Asteggiante, Lucía; Belew, Ashton T; Clements, Virginia K et al. (2018) Differential Content of Proteins, mRNAs, and miRNAs Suggests that MDSC and Their Exosomes May Mediate Distinct Immune Suppressive Functions. J Proteome Res 17:486-498
Singh, Rajesh K; Kazansky, Yaniv; Wathieu, Donald et al. (2017) Hydrophobic Patch of Ubiquitin is Important for its Optimal Activation by Ubiquitin Activating Enzyme E1. Anal Chem 89:7852-7860
Bronte, Vincenzo; Brandau, Sven; Chen, Shu-Hsia et al. (2016) Recommendations for myeloid-derived suppressor cell nomenclature and characterization standards. Nat Commun 7:12150
Choksawangkarn, Waeowalee; Graham, Lauren M; Burke, Meghan et al. (2016) Peptide-based systems analysis of inflammation induced myeloid-derived suppressor cells reveals diverse signaling pathways. Proteomics 16:1881-8
Kim, Yeji; Edwards, Nathan; Fenselau, Catherine (2016) Extracellular vesicle proteomes reflect developmental phases of Bacillus subtilis. Clin Proteomics 13:6
Castañeda, Carlos A; Dixon, Emma K; Walker, Olivier et al. (2016) Linkage via K27 Bestows Ubiquitin Chains with Unique Properties among Polyubiquitins. Structure 24:423-36
Lee, Amanda E; Geis-Asteggiante, Lucia; Dixon, Emma K et al. (2016) Preparing to read the ubiquitin code: characterization of ubiquitin trimers by top-down mass spectrometry. J Mass Spectrom 51:315-21
Geis-Asteggiante, Lucía; Ostrand-Rosenberg, Suzanne; Fenselau, Catherine et al. (2016) Evaluation of Spectral Counting for Relative Quantitation of Proteoforms in Top-Down Proteomics. Anal Chem 88:10900-10907

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