The broad goal of this competing renewal focuses on glycosaminoglycan separation, purification and complete chemical characterization as it pertains to heparin and HS complexation to various inflammatory chemokines. Methods will be developed to separate, purify and identify glycosaminoglycan (GAG) binders of three specific Rheumatoid Arthritis associated chemokines using chromatography, ion mobility mass spectrometry (IM-MS) and CID-MS. Complexation of GAG binders to their chemokine counterparts will be investigated by MS with the monomer, dimer and tetramer assemblies measured using IM-MS. These studies will be paired with the compositional analysis of GAG in sera from women with Rheumatoid Arthritis in the pre- and postmenopausal age range to determine health benefits of HRT and its correlation with GAG composition. The three specific aims are:
Specific Aim 1 : GAG composition in Human Serum, Specific Aim 2: Chromatographic preparation, separation, identification and complexation of GAG libraries with Chemokines, and Specific Aim 3: Ion Mobility MS and X-ray crystallography of Chemokine and GAG binders. The major areas proposed are interwoven in such a way that methods development, biotechnology and application to biologically relevant systems are the cohesive elements that bind the proposed research together.
This research will develop methods to separate, purify and identify glycosaminoglycan (GAG) binders of various inflammatory chemokines using chromatography and ion mobility mass spectrometry (IMS). Complexation of these binders to their chemokine counterparts will be investigated by MS and multimeric assemblies measured using theoretical and experimental collision cross sections via IMS. These studies will be paired with the compositional analysis of GAG in serum from Rheumatoid Arthritis women in the pre- and postmenopausal age range to determine health benefits of HRT and its correlation with GAG composition.
|Sogi, Kimberly M; Holsclaw, Cynthia M; Fragiadakis, Gabriela K et al. (2016) Biosynthesis and Regulation of Sulfomenaquinone, a Metabolite Associated with Virulence in Mycobacterium tuberculosis. ACS Infect Dis 2:800-806|
|Leary, Julie A; Miller, Rebecca L; Wei, Wei et al. (2015) Composition, sequencing and ion mobility mass spectrometry of heparan sulfate-like octasaccharide isomers differing in glucuronic and iduronic acid content. Eur J Mass Spectrom (Chichester) 21:245-54|
|Nithianantham, Stanley; Le, Sinh; Seto, Elbert et al. (2015) Tubulin cofactors and Arl2 are cage-like chaperones that regulate the soluble ??-tubulin pool for microtubule dynamics. Elife 4:|
|Sabol, Jenny K; Wei, Wei; López-Hoyos, Marcos et al. (2014) Heparan sulfate differences in rheumatoid arthritis versus healthy sera. Matrix Biol 40:54-61|
|Seo, Youjin; Andaya, Armann; Bleiholder, Christian et al. (2013) Differentiation of CC vs CXC chemokine dimers with GAG octasaccharide binding partners: an ion mobility mass spectrometry approach. J Am Chem Soc 135:4325-32|
|Wei, Wei; Miller, Rebecca L; Leary, Julie A (2013) Method development and analysis of free HS and HS in proteoglycans from pre- and postmenopausal women: evidence for biosynthetic pathway changes in sulfotransferase and sulfatase enzymes. Anal Chem 85:5917-23|
|Seo, Youjin; Andaya, Armann; Leary, Julie A (2012) Preparation, separation, and conformational analysis of differentially sulfated heparin octasaccharide isomers using ion mobility mass spectrometry. Anal Chem 84:2416-23|
|Ninonuevo, Milady R; Leary, Julie A (2012) Ion mobility mass spectrometry coupled with rapid protein threading predictor structure prediction and collision-induced dissociation for probing chemokine conformation and stability. Anal Chem 84:3208-14|
|Seo, Youjin; Schenauer, Matthew R; Leary, Julie A (2011) Biologically Relevant Metal-Cation Binding Induces Conformational Changes in Heparin Oligosaccharides as Measured by Ion Mobility Mass Spectrometry. Int J Mass Spectrom 303:191-198|
|Wei, Wei; Ninonuevo, Milady R; Sharma, Anish et al. (2011) A comprehensive compositional analysis of heparin/heparan sulfate-derived disaccharides from human serum. Anal Chem 83:3703-8|
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