The goal of this work is to correlate heparan sulfate structure with angiogenic function in vascular tissue. Both normal and pathological angiogenesis are mediated through growth factor stimuli that depend on the structures of cell surface and extracellular matrix heparan sulfate (HS) chains for receptor activation. HS serves as both a spatial and temporal regulator of growth factor activity during angiogenesis. The diversity of HS biological activities arises through their non-template driven biosynthesis. A series of modifying enzymes act upon nascent HS chains to produce mature molecules with characteristic domains of high and low sulfation. Chain lengths and degree of sulfation are heterogeneous and reflect the responses of cells to their growth environment. Variation in expression of HS chain structure is a mechanism whereby cells modulate their responses to growth factor stimuli. In this sense, the diversity of HS chains on cell surfaces and secreted proteins is a means of elaborating the functions of a limited array of growth factors and growth factor receptors. In vascular tissue, homeostasais and mitogenesis are controlled through growth factor signalling cascades. Fibroblast growth factors (FGFs), and their receptors bind HS chains on the cell surface and the extracellular matrix. At the present time, it is clear that both high and low affinity HS domains exist and play important roles in modulating angiogenic responses. Although it is also clear that such subsequences may either potentiate or inhibit growth factor, depending on the context, there is little information concerning their structures. In order to address these questions, new methods will be developed in Aim 1 to enable sequencing of HS compatible with on-line liquid chromatography-tandem mass spectrometry.
In Aim 2, libraries of organ-specific HS will be prepared using two dimensional chromatography. The structures will be determined using on-line tandem mass spectrometry and correlated with function using cell free growth factor binding and cellular mitogenesis assays.
Aim 3 is to determine the manner in which extracellular enzymes (mammalian endo- sulfatases and heparanase) remodel HS chains in biological systems.
In vascular tissue, both normal and pathological cell growth related to angiogenesis is regulated through growth factor signaling cascades. Cells modulate their responses to growth factor signaling by altering the structures of heparan sulfate chains expressed on their surfaces and secreted into the extracellular matrix. This research will explore the structure-function relationships of vascular heparan sulfates in order to inform efforts to develop protein binding microarrays, drugs and therapeutics.
|Hu, Han; Khatri, Kshitij; Zaia, Joseph (2017) Algorithms and design strategies towards automated glycoproteomics analysis. Mass Spectrom Rev 36:475-498|
|Reine, Trine Marita; Kolseth, Ingrid Benedicte Moss; Meen, Astri Jeanette et al. (2015) Effects of restoring normoglycemia in type 1 diabetes on inflammatory profile and renal extracellular matrix structure after simultaneous pancreas and kidney transplantation. Diabetes Res Clin Pract 107:46-53|
|Huang, Yu; Mao, Yang; Zong, Chengli et al. (2015) Discovery of a heparan sulfate 3-O-sulfation specific peeling reaction. Anal Chem 87:592-600|
|Mao, Yang; Huang, Yu; Buczek-Thomas, Jo Ann et al. (2014) A liquid chromatography-mass spectrometry-based approach to characterize the substrate specificity of mammalian heparanase. J Biol Chem 289:34141-51|
|Turiák, Lilla; Shao, Chun; Meng, Le et al. (2014) Workflow for combined proteomics and glycomics profiling from histological tissues. Anal Chem 86:9670-8|
|Huang, Yu; Mao, Yang; Buczek-Thomas, Jo Ann et al. (2014) Oligosaccharide substrate preferences of human extracellular sulfatase Sulf2 using liquid chromatography-mass spectrometry based glycomics approaches. PLoS One 9:e105143|
|Ramachandra, Rashmi; Namburi, Ramesh B; Ortega-Martinez, Olga et al. (2014) Brittlestars contain highly sulfated chondroitin sulfates/dermatan sulfates that promote fibroblast growth factor 2-induced cell signaling. Glycobiology 24:195-207|
|Hu, Han; Huang, Yu; Mao, Yang et al. (2014) A computational framework for heparan sulfate sequencing using high-resolution tandem mass spectra. Mol Cell Proteomics 13:2490-502|
|Shi, Xiaofeng; Shao, Chun; Mao, Yang et al. (2013) LC-MS and LC-MS/MS studies of incorporation of 34SO3 into glycosaminoglycan chains by sulfotransferases. Glycobiology 23:969-79|
|Nugent, M A; Zaia, J; Spencer, J L (2013) Heparan sulfate-protein binding specificity. Biochemistry (Mosc) 78:726-35|
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