Thisprojectwilldevelopanautomatedcapillaryelectrophoresis-tandemmassspectrometryplatformforthe structuralanalysisofmixturesofglycosaminoglycanoligomersthatareisolatedinproteinbindingassays.This classofcarbohydratescanbefoundonthesurfaceofallanimalcells,andplayakeyroleinmanyimportant biologicalprocesses.Glycosaminoglycansarelinear,polysulfatedcopolymers,typicallywitharepeating disaccharideunitcomposedofanN-acetyl-hexosamineandglucuronicacid,andcanbemorethan200repeat unitslong.Modificationscanoccuranywherealongthechain,andtheseincludeO-sulfation,N- deacetylation/N-sulfation,andepimerizationoftheuronicacid.Theseareaparticularlychallengingclassof compoundstocharacterizeduetotheirheterogeneousstructures,anoutcomeoftheirnon-template biosynthesis.Nevertheless,withinthelongglycanchainsareshortregionsofdefinedmodificationthattarget specificproteinswithremarkablespecificity.Theseglycan-proteininteractionsarethebasisforthebiological activityoftheglycosaminoglycan.Thereisacriticalneedfortoolstofindtheseorganizedregionswiththelong glycanchain,andtodeterminetheirpatternofmodification.Afewspecializedlaboratorieshavedeveloped massspectrometrymethodsforthestructuralanalysisofglycosaminoglycans,butthesehavenotfoundtheir wayintobiologicallaboratoriesatlarge.Thegoalofthisprojectistoproduceaplatformwithahighdegreeof automationsothatitcanbedeployedinthelaboratoriesofbiologyresearchers.Theaccessibilityofthese advancedmassspectrometrymethodswillresultfrominnovationsinhardware,software,andmethods development.

Public Health Relevance

Glycosaminoglycansaresulfatedcarbohydratesthatplayacentralroleinawiderangeofnormaland pathologicalprocessesinanimalcells.Currently,thereisnotechnologyforthestructuralanalysisofmixtures ofthisclassofcarbohydrates,whichlimitsthecapabilitytostudytheirbiologicalactivitythroughproteinbinding assays.Thisproposalpresentsaplanforanautomatedplatformforanalyzingcomplexmixturesof glycosaminoglycans,utilizingcapillaryelectrophoresisandtandemmassspectrometry,thatcanbedeployed intothelaboratoriesofbiologicalandbiomedicalresearchers.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01CA231074-02
Application #
9753175
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Krueger, Karl E
Project Start
2018-08-01
Project End
2021-07-31
Budget Start
2019-08-01
Budget End
2020-07-31
Support Year
2
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of Georgia
Department
Chemistry
Type
Graduate Schools
DUNS #
004315578
City
Athens
State
GA
Country
United States
Zip Code
30602