Heparan sulfate (HS) is an essential glycan that is present in large quantities on the cell surface and in the extracellular matrix. HS participates in a variety of physiological and pathophysiological functions, including blood coagulation, inflammatory response and cell differentiation. HS is a highly sulfated polysaccharide. Heparin, a special form of HS, is a commonly used anticoagulant drug. The wide range of biological functions of HS attracts considerable interest in exploiting heparin or heparin-like molecules for the development of anticancer and antiviral drugs. The uniquely distributed sulfation pattern of HS is believed to regulate its functional specificity. However, chemical synthesis of HS, especially those larger than hexasaccharides, is extremely difficult. Using HS biosynthetic enzymes, our labs can produce an array of HS with unique sulfation patterns and functions. Our success has proved the feasibility of conducting enzyme-based synthesis of HS with unique biological activities. The long term aim of this project is to develop a method to synthesize HS with high structural precision. Our hypothesis is that the distribution of N- sulfoglucosamine residues determines the susceptibility of all subsequent modifications during heparan sulfate biosynthesis, including epimerization, 2-O-sulfation, 6-O- sulfation, and 3-O-sulfation. In this proposal, we will synthesize structurally defined oligosaccharides carrying N-sulfoglucosamine residues using glycosyl transferases. We will then determine the substrate specificities of a variety of HS biosynthetic enzymes with the aim of precisely positioning the 6-O-sulfo and 3-O-sulfo groups as well as 2-O- sulfo iduronic acid within oligosaccharides. Finally, we plan to utilize this method to investigate novel anticoagulant HS structures. The success of this project will lead a comprehensive new approach to investigate the structure and function relationship of HS, potentially leading to the development of novel HS-based therapeutic reagents. Public Health Relevance: Heparan sulfate (HS) is a highly sulfated polysaccharide. Heparin, a special form of HS, is a commonly used anticoagulant drug. The wide range of biological functions of HS attracts considerable interest in exploiting heparin or heparin-like molecules for the development of anticancer and antiviral drugs. The uniquely distributed sulfation pattern of HS is believed to regulate its functional specificity. The long term aim of this project is to develop a method to synthesize HS with high structural precision. The success of this project will lead a comprehensive new approach to investigate the structure and function relationship of HS, potentially leading to the development of novel HS-based therapeutic reagents.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL094463-02
Application #
7767769
Study Section
Intercellular Interactions (ICI)
Program Officer
Sarkar, Rita
Project Start
2009-02-13
Project End
2013-01-31
Budget Start
2010-02-01
Budget End
2011-01-31
Support Year
2
Fiscal Year
2010
Total Cost
$423,870
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Pellock, Samuel J; Walton, William G; Biernat, Kristen A et al. (2018) Three structurally and functionally distinct ?-glucuronidases from the human gut microbe Bacteroides uniformis. J Biol Chem 293:18559-18573
Xu, Ding; Arnold, Katelyn; Liu, Jian (2018) Using structurally defined oligosaccharides to understand the interactions between proteins and heparan sulfate. Curr Opin Struct Biol 50:155-161
Stancanelli, Eduardo; Elli, Stefano; Hsieh, Po-Hung et al. (2018) Recognition and Conformational Properties of an Alternative Antithrombin Binding Sequence Obtained by Chemoenzymatic Synthesis. Chembiochem :
Xiao, Yiming; Li, Miaomiao; Larocque, Rinzhi et al. (2018) Dimerization interface of osteoprotegerin revealed by hydrogen-deuterium exchange mass spectrometry. J Biol Chem 293:17523-17535
Lin, Yi-Pin; Li, Lingyun; Zhang, Fuming et al. (2017) Borrelia burgdorferi glycosaminoglycan-binding proteins: a potential target for new therapeutics against Lyme disease. Microbiology 163:1759-1766
Wang, Zhangjie; Hsieh, Po-Hung; Xu, Yongmei et al. (2017) Synthesis of 3-O-Sulfated Oligosaccharides to Understand the Relationship between Structures and Functions of Heparan Sulfate. J Am Chem Soc :
Xu, Yongmei; Moon, Andrea F; Xu, Shuqin et al. (2017) Structure Based Substrate Specificity Analysis of Heparan Sulfate 6-O-Sulfotransferases. ACS Chem Biol 12:73-82
Meneghetti, Maria Cecília Zorél; Gesteira Ferreira, Tarsis; Tashima, Alexandre Keiji et al. (2017) Insights into the role of 3-O-sulfotransferase in heparan sulfate biosynthesis. Org Biomol Chem 15:6792-6799
Xu, Yongmei; Chandarajoti, Kasemsiri; Zhang, Xing et al. (2017) Synthetic oligosaccharides can replace animal-sourced low-molecular weight heparins. Sci Transl Med 9:
Schultz, Victor; Suflita, Mathew; Liu, Xinyue et al. (2017) Heparan Sulfate Domains Required for Fibroblast Growth Factor 1 and 2 Signaling through Fibroblast Growth Factor Receptor 1c. J Biol Chem 292:2495-2509

Showing the most recent 10 out of 85 publications