Post-biosynthetic remodeling of heparan sulfate

Summary

Principal Investigator: Joseph Zaia
Abstract: DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: 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.
Funding Period: 2010-01-18 - 2014-12-31
more information: NIH RePORT

Top Publications

  1. pmc Mass spectrometry and the emerging field of glycomics
    Joseph Zaia
    Deptartment of Biochemistry, Boston University, 670 Albany Street, Boston, MA 02118, USA
    Chem Biol 15:881-92. 2008
  2. pmc Mass spectrometric method for determining the uronic acid epimerization in heparan sulfate disaccharides generated using nitrous acid
    Vanessa Leah Gill
    Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts, United States
    Anal Chem 84:7539-46. 2012
  3. pmc Top-down approach for the direct characterization of low molecular weight heparins using LC-FT-MS
    Lingyun Li
    Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, New York 12180 3590, United States
    Anal Chem 84:8822-9. 2012
  4. pmc GlycReSoft: a software package for automated recognition of glycans from LC/MS data
    Evan Maxwell
    Bioinformatics Program, Center for Biomedical Mass Spectrometry, Department, of Biochemistry, Boston University, Boston, Massachusetts, USA
    PLoS ONE 7:e45474. 2012
  5. pmc Disaccharide analysis of glycosaminoglycans using hydrophilic interaction chromatography and mass spectrometry
    Vanessa Leah Gill
    Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts 02118, USA
    Anal Chem 85:1138-45. 2013
  6. pmc Glycosaminoglycan glycomics using mass spectrometry
    Joseph Zaia
    Center for Biomedical Mass Spectrometry, Department of Biochemistry, Boston University, Boston, Massachusetts 02118, USA
    Mol Cell Proteomics 12:885-92. 2013
  7. pmc Capillary electrophoresis-mass spectrometry of carbohydrates
    Joseph Zaia
    Department of Biochemistry, Center for Biomedical Mass Spectrometry, Boston University Medial Campus, Boston, MA, USA
    Methods Mol Biol 984:13-25. 2013
  8. pmc Comparative glycomics of leukocyte glycosaminoglycans
    Chun Shao
    Department of Biochemistry, Boston University School of Medicine, Boston University Medical Campus, MA 02118, USA
    FEBS J 280:2447-61. 2013
  9. pmc LC-MS and LC-MS/MS studies of incorporation of 34SO3 into glycosaminoglycan chains by sulfotransferases
    Xiaofeng Shi
    Department of Biochemistry and Center for Biomedical Mass Spectrometry, Boston University School of Medicine, Boston, MA 02118, USA
    Glycobiology 23:969-79. 2013
  10. pmc Heparan sulfate-protein binding specificity
    M A Nugent
    Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118, USA
    Biochemistry (Mosc) 78:726-35. 2013

Detail Information

Publications28

  1. pmc Mass spectrometry and the emerging field of glycomics
    Joseph Zaia
    Deptartment of Biochemistry, Boston University, 670 Albany Street, Boston, MA 02118, USA
    Chem Biol 15:881-92. 2008
    ..Mass spectrometry (MS) is emerging as an enabling technology in the field of glycomics. This review summarizes recent developments in mass spectrometric analysis methods for protein-based glycomics and glycoproteomics workflows...
  2. pmc Mass spectrometric method for determining the uronic acid epimerization in heparan sulfate disaccharides generated using nitrous acid
    Vanessa Leah Gill
    Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts, United States
    Anal Chem 84:7539-46. 2012
    ..The results are comparable to those expected for benchmark HS and heparin samples. The data demonstrate the utility of PGC-MS for quantification of HS nitrous acid depolymerization products for structural analysis of HS and heparin...
  3. pmc Top-down approach for the direct characterization of low molecular weight heparins using LC-FT-MS
    Lingyun Li
    Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, Troy, New York 12180 3590, United States
    Anal Chem 84:8822-9. 2012
    ..Bioinformatics software (GlycReSoft 1.0) was used to automatically assign structures within 5-ppm mass accuracy...
  4. pmc GlycReSoft: a software package for automated recognition of glycans from LC/MS data
    Evan Maxwell
    Bioinformatics Program, Center for Biomedical Mass Spectrometry, Department, of Biochemistry, Boston University, Boston, Massachusetts, USA
    PLoS ONE 7:e45474. 2012
    ..We demonstrate this tool, GlycReSoft, using an LC/MS dataset on tissue derived heparan sulfate oligosaccharides. The software, code and a test data set are publically archived under an open source license...
  5. pmc Disaccharide analysis of glycosaminoglycans using hydrophilic interaction chromatography and mass spectrometry
    Vanessa Leah Gill
    Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts 02118, USA
    Anal Chem 85:1138-45. 2013
    ..This method enables biomedical researchers to determine complete disaccharide profiles on GAG samples using a single LC-MS platform...
  6. pmc Glycosaminoglycan glycomics using mass spectrometry
    Joseph Zaia
    Center for Biomedical Mass Spectrometry, Department of Biochemistry, Boston University, Boston, Massachusetts 02118, USA
    Mol Cell Proteomics 12:885-92. 2013
    ..This review summarizes progress on mass-spectrometry-based GAG glycomics methods. The areas covered include disaccharide analysis, oligosaccharide profiling, and tandem mass spectrometric sequencing...
  7. pmc Capillary electrophoresis-mass spectrometry of carbohydrates
    Joseph Zaia
    Department of Biochemistry, Center for Biomedical Mass Spectrometry, Boston University Medial Campus, Boston, MA, USA
    Methods Mol Biol 984:13-25. 2013
    ..This chapter summarizes applications of CS/MS to analysis of carbohydrates, glycoproteins, and glycopeptides that have appeared since 2008. Readers are referred to recent comprehensive reviews covering earlier publications...
  8. pmc Comparative glycomics of leukocyte glycosaminoglycans
    Chun Shao
    Department of Biochemistry, Boston University School of Medicine, Boston University Medical Campus, MA 02118, USA
    FEBS J 280:2447-61. 2013
    ..This information establishes the range of GAG structures expressed on normal leukocytes and is necessary for subsequent inquiry into disease conditions...
  9. pmc LC-MS and LC-MS/MS studies of incorporation of 34SO3 into glycosaminoglycan chains by sulfotransferases
    Xiaofeng Shi
    Department of Biochemistry and Center for Biomedical Mass Spectrometry, Boston University School of Medicine, Boston, MA 02118, USA
    Glycobiology 23:969-79. 2013
    ..Contrary to the traditional view of HS biosynthesis processes, NDST1 also showed activity on O-sulfated GlcNAc residues. ..
  10. pmc Heparan sulfate-protein binding specificity
    M A Nugent
    Department of Biochemistry, Boston University School of Medicine, Boston, MA 02118, USA
    Biochemistry (Mosc) 78:726-35. 2013
    ....
  11. pmc Mass spectral profiling of glycosaminoglycans from histological tissue surfaces
    Chun Shao
    Center for Biomedical Mass Spectrometry, Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts 02118, United States
    Anal Chem 85:10984-91. 2013
    ..Higher HS abundances and lower average sulfation level of HS were detected in glioblastoma (GBM, WHO grade IV) slides compared to astrocytoma (WHO grade II) slides. ..
  12. pmc De novo sequencing of heparan sulfate oligosaccharides by electron-activated dissociation
    Yu Huang
    Mass Spectrometry Resource, Department of Biochemistry, Boston University School of Medicine, 670 Albany Street, Suite 504, Boston, Massachusetts 02118, United States
    Anal Chem 85:11979-86. 2013
    ..These results demonstrate the potential of EDD and NETD as sensitive analytical tools for detailed, high-throughput, de novo structural analyses of highly sulfated GAGs. ..
  13. pmc Brittlestars contain highly sulfated chondroitin sulfates/dermatan sulfates that promote fibroblast growth factor 2-induced cell signaling
    Rashmi Ramachandra
    Department of Medical Biochemistry and Microbiology, The Biomedical Center, Uppsala University, Box 582, SE 751 23 Uppsala, Sweden
    Glycobiology 24:195-207. 2014
    ..These findings point to a potential biological role for the highly sulfated invertebrate GAGs, similar to those ascribed to HS in vertebrates. ..
  14. pmc Heparan sulfate 6-O-endosulfatases (Sulfs) coordinate the Wnt signaling pathways to regulate myoblast fusion during skeletal muscle regeneration
    Thanh H Tran
    The Pulmonary Center, Department of Medicine, Boston University School of Medicine, Boston, Massachusetts 02118, USA
    J Biol Chem 287:32651-64. 2012
    ..Together, our findings uncover a critical role of Sulfs in myoblast fusion by promoting antagonizing canonical Wnt signaling activities against the noncanonical Wnt pathway during skeletal muscle regeneration...
  15. pmc Tandem mass spectrometry of heparan sulfate negative ions: sulfate loss patterns and chemical modification methods for improvement of product ion profiles
    Xiaofeng Shi
    Department of Biochemistry and Center for Biomedical Mass Spectrometry, Boston University School of Medicine, Boston, MA 02118, USA
    J Am Soc Mass Spectrom 23:1498-511. 2012
    ..These modifications effectively reduced the sulfate density on the HS oligosaccharides and generated considerably more backbone dissociation using on-line LC/tandem MS...
  16. pmc Improved hydrophilic interaction chromatography LC/MS of heparinoids using a chip with postcolumn makeup flow
    Gregory O Staples
    Mass Spectrometry Resource, Department of Biochemistry, Boston University School of Medicine, 670 Albany Street, Boston, Massachusetts 02118, USA
    Anal Chem 82:516-22. 2010
    ..It is expected that this technology will enable quantitative, comparative glycomics profiling of extended GAG oligosaccharide domains of functional interest...
  17. pmc Comparative glycomics using a tetraplex stable-isotope coded tag
    Michael J Bowman
    Boston University School of Medicine, Department of Biochemistry, Center for Biomedical Mass Spectrometry, Boston, Massachusetts 02118, USA
    Anal Chem 82:3023-31. 2010
    ..The data demonstrate the value of the tetraplex stable isotope tagging approach for producing high-quality glycomics compositional profiling and fine structural analysis...
  18. pmc Screening for anticoagulant heparan sulfate octasaccharides and fine structure characterization using tandem mass spectrometry
    Hicham Naimy
    Department of Biochemistry, Boston University School of Medicine, 670 Albany Street, Boston, Massachusetts 02118, USA
    Biochemistry 49:3743-52. 2010
    ..This approach confirmed isomeric enrichment of these compositions and, most importantly, produces ions diagnostic of their biological activity...
  19. pmc Targeted analysis of glycomics liquid chromatography/mass spectrometry data
    Jonathan M Dreyfuss
    Graduate Program in Bioinformatics, Boston University, Boston, MA 02218, USA
    Anal Bioanal Chem 399:727-35. 2011
    ..The capabilities of the tool were demonstrated using a set of HILIC LC/MS data on organ-specific heparan sulfates...
  20. pmc Glycomics analysis of mammalian heparan sulfates modified by the human extracellular sulfatase HSulf2
    Gregory O Staples
    Department of Biochemistry, Center for Biomedical Mass Spectrometry, Boston University School of Medicine, Boston, Massachusetts, United States of America
    PLoS ONE 6:e16689. 2011
    ..Therefore, the present work aims to extend the understanding of substrate specificities of HSulf2 using in vitro experiments to compare HSulf2 activities on HS from different organ tissues...
  21. pmc Highly sulfated nonreducing end-derived heparan sulfate domains bind fibroblast growth factor-2 with high affinity and are enriched in biologically active fractions
    Hicham Naimy
    Department of Biochemistry, Center for Biomedical Mass Spectrometry, Boston University School of Medicine, Boston, Massachusetts 02118, USA
    J Biol Chem 286:19311-9. 2011
    ..The results suggest a role of the NRE of HS in FGF2 signaling and favor the formation of the symmetrical architecture on short NS domains...
  22. pmc WT1-dependent sulfatase expression maintains the normal glomerular filtration barrier
    Valérie A Schumacher
    Department of Medicine, Children s Hospital Boston and Department of Pediatrics, Harvard Medical School, Boston, Massachusetts 02115, USA
    J Am Soc Nephrol 22:1286-96. 2011
    ....
  23. pmc Improved liquid chromatography-MS/MS of heparan sulfate oligosaccharides via chip-based pulsed makeup flow
    Yu Huang
    Center for Biomedical Mass Spectrometry, Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts 02118, United States
    Anal Chem 83:8222-9. 2011
    ..Tandem MS of these supercharged precursor ions showed decreased abundances of product ions from sulfate losses yet more abundant product ions from backbone cleavages...
  24. pmc Dermatan sulfate is involved in the tumorigenic properties of esophagus squamous cell carcinoma
    Martin A Thelin
    Department of Experimental Medical Science, Biomedical Center D12, Lund University, Lund, Sweden
    Cancer Res 72:1943-52. 2012
    ..Our findings show that IdoA in DS influences tumorigenesis by affecting cancer cell behavior. Therefore, downregulation of IdoA by DS-epi1 inhibitors may represent a new anticancer therapy...
  25. pmc Effective use of mass spectrometry for glycan and glycopeptide structural analysis
    Nancy Leymarie
    Department of Biochemistry, Center for Biomedical Mass Spectrometry, Boston University Medical Campus, Boston, Massachusetts 02118, USA
    Anal Chem 84:3040-8. 2012
    ..This technology enables precise characterization of recombinant glycoproteins in the pharmaceutical industry and academic biomedicine...
  26. pmc A small molecule glycosaminoglycan mimetic blocks Plasmodium invasion of the mosquito midgut
    Derrick K Mathias
    W Harry Feinstone Department of Molecular Microbiology and Immunology, Malaria Research Institute, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, United States of America
    PLoS Pathog 9:e1003757. 2013
    ..We envision that such compounds when used as partner drugs with current antimalarial regimens and with RTS,S vaccine delivery could prevent the transmission of drug-resistant and vaccine-breakthrough strains. ..