Heparan sulfates (HS) are abundantly expressed on the cell surface and in the extracellular matrix as proteoglycans with differential sulfation pattern, chain length and number of chains leading to vast structural features. Each chain interacts simultaneously with numerous proteins including a variety of signaling molecules, morphogens, receptors, etc and orchestrates various biological functions. Modulation of their biosynthetic pathways has begun to shed light on their role in such model organisms as zebra fish, c. elegans, drosophila, etc. However, the lack of efficient synthetic approaches coupled with a small amount of HS chains isolated from animal models has limited our knowledge of how these molecules regulate various physiological and pathological processes at the molecular level. The central hypothesis of this proposal is that multiple oligosaccharide sequences with conserved critical groups may elucidate the same biological response or that a given oligosaccharide sequence with a set of critical groups may elucidate more than one biological response. Determining those critical groups and their spatial distance would be sufficient to determine the rules that dictate HS-protein interactions. Development of a general enzymatic approach for the rapid assembly of HS of various sizes should allow us to test our hypothesis and to delineate the rules that dictate HS-protein interactions. We propose to construct the oligosaccharide libraries and screen their ability to form signaling complex with various growth factors and receptors that play a central role in heart, lung and brain development. With the aid of stable isotopes, we will then develop novel structural biology approaches to elucidate the critical groups that attribute a biological function. The knowledge gained in synthesizing ATIII binding sites will be used in designing diverse size-based HS libraries that should be the key to defining all interacting proteins and elucidating the chemical rules by which such interactions take place. This proposed study should establish a firm foundation for the glycobiology of proteoglycans.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM075168-04
Application #
7884258
Study Section
Special Emphasis Panel (ZRG1-ICI-G (01))
Program Officer
Marino, Pamela
Project Start
2007-09-26
Project End
2012-06-30
Budget Start
2010-07-01
Budget End
2011-06-30
Support Year
4
Fiscal Year
2010
Total Cost
$283,091
Indirect Cost
Name
University of Utah
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
009095365
City
Salt Lake City
State
UT
Country
United States
Zip Code
84112
Joice, April; Raman, Karthik; Mencio, Caitlin et al. (2015) Enzymatic synthesis of heparan sulfate and heparin. Methods Mol Biol 1229:11-9
Babu, Ponnusamy; Victor, Xylophone V; Raman, Karthik et al. (2015) A rapid, nonradioactive assay for measuring heparan sulfate C-5 epimerase activity using hydrogen/deuterium exchange-mass spectrometry. Methods Mol Biol 1229:209-19
Raman, Karthik; Kuberan, Balagurunathan; Arungundram, Sailaja (2015) Chemical modification of heparin and heparosan. Methods Mol Biol 1229:31-6
Kalita, Mausam; Quintero, Maritza V; Raman, Karthik et al. (2015) Synthesis and biomedical applications of xylosides. Methods Mol Biol 1229:517-28
Brown, Spencer; Kuberan, Balagurunathan (2015) Production of size-defined heparosan, heparan sulfate, and heparin oligosaccharides by enzymatic depolymerization. Methods Mol Biol 1229:21-9
Mencio, Caitlin; Garud, Dinesh R; Kuberan, Balagurunathan et al. (2015) Synthesis of selective inhibitors of heparan sulfate and chondroitin sulfate proteoglycan biosynthesis. Methods Mol Biol 1229:69-78
Victor, Xylophone V; Tran, Vy M; Kuberan, Balagurunathan et al. (2015) Preparation of isotope-enriched heparan sulfate precursors for structural biology studies. Methods Mol Biol 1229:43-8
Mencio, Caitlin; Swarup, Vimal P; Soliai, Marcus et al. (2015) Synthesis of sulfur isotope-labeled sulfate donor, 3'-phosphoadenosine-5'-phosphosulfate, for studying glycosaminoglycan functions. Methods Mol Biol 1229:37-42
Tran, Vy M; Kuberan, Balagurunathan (2014) Synthesis of fluorophore-tagged xylosides that prime glycosaminoglycan chains. Bioconjug Chem 25:262-8
Kalita, Mausam; Balivada, Sivasai; Swarup, Vimal Paritosh et al. (2014) A nanosensor for ultrasensitive detection of oversulfated chondroitin sulfate contaminant in heparin. J Am Chem Soc 136:554-7

Showing the most recent 10 out of 30 publications