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.

Public Health Relevance

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.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM047356-20A1
Application #
8513593
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Edmonds, Charles G
Project Start
1992-05-01
Project End
2017-05-31
Budget Start
2013-09-30
Budget End
2014-05-31
Support Year
20
Fiscal Year
2013
Total Cost
$252,189
Indirect Cost
$75,526
Name
University of California Davis
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
047120084
City
Davis
State
CA
Country
United States
Zip Code
95618
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
Holley, Rebecca J; Deligny, Audrey; Wei, Wei et al. (2011) Mucopolysaccharidosis type I, unique structure of accumulated heparan sulfate and increased N-sulfotransferase activity in mice lacking ?-l-iduronidase. J Biol Chem 286:37515-24
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
Jen, Connie H; Leary, Julie A (2010) A competitive binding study of chemokine, sulfated receptor, and glycosaminoglycan interactions by nano-electrospray ionization mass spectrometry. Anal Biochem 407:134-40
Schenauer, Matthew R; Leary, Julie A (2009) An Ion Mobility-Mass Spectrometry Investigation of Monocyte Chemoattractant Protein-1. Int J Mass Spectrom 287:70-76
Meissen, John K; Sweeney, Matthew D; Girardi, Matthew et al. (2009) Differentiation of 3-O-sulfated heparin disaccharide isomers: identification of structural aspects of the heparin CCL2 binding motif. J Am Soc Mass Spectrom 20:652-7

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