University of Illinois Urbana-Champaign

Analysis of the Structural Units of Sulfated Glycosaminoglycans by High Performance Ion Exchange Chromatography

Delaney, Stephen Ralph

This item is only available for download by members of the University of Illinois community. Students, faculty, and staff at the U of I may log in with your NetID and password to view the item. If you are trying to access an Illinois-restricted dissertation or thesis, you can request a copy through your library's Inter-Library Loan office or purchase a copy directly from ProQuest.

Permalink

https://hdl.handle.net/2142/67417

Description

Title
Analysis of the Structural Units of Sulfated Glycosaminoglycans by High Performance Ion Exchange Chromatography
Author(s)
Delaney, Stephen Ralph
Issue Date
1981
Department of Study
Biochemistry
Discipline
Biochemistry
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Chemistry, Biochemistry
Language
eng
Abstract
  • The structural units of heparin produced by nitrous acid deamination were stoichiometrically reduced with NaB('3)H(,4) and analyzed by high performance ion exchange chromatography to obtain resolution of the four primary disaccharide units of heparin; 4-O-((beta)-D-glucopyranosyluronic acid)-6-O-sulfo-2,5-anhydro-D-mannitol (GMS), two-L-iduronic acid-containing monosulfated disaccharides, 4-O-((alpha)-L-idopyranosyluronic acid)-6-O-sulfo-2,5-anhydro-D-mannitol (IMS) and 4-O-(2-O-sulfo-(alpha)-L-idopyranosyluronic acid)-2,5-anhydro-D-mannitol (ISM), and a single disulfated disaccharide, 4-O-(2-O-sulfo-(alpha)-L-idopyranosyluronic acid)-6-O-sulfo-2,5-anhydro-D-mannitol (ISMS). The capacity to separate tetrasaccharides or higher oligosaccharides was demonstrated using chondroitin sulfate oligosaccharides which could be structurally characterized in terms of their chondroitinase ABC digestion products. The high performance ion exchange chromatography technique has been used in the analysis of the oligosaccharide composition of a series of heparins fractionated by Dowex anion exchange chromatography and in the isolation and characterization of unidentified deamination oligosaccharides which appear in smaller amounts in the hplc profiles.
  • The analyses of the Dowex-fractionated heparins showed that the monosulfated disaccharide ISM and other uncharacterized monosulfated oligosaccharides are the major oligosaccharide units in Dowex fractions which elute in low salt. In fractions eluting at high ionic strength, the disulfated disaccharide, ISMS and the trisulfated tetrasaccharide involved in the specific binding of heparin to antithrombin III binding site are the main structural units in the deamination products of these fractions. The increase in the antithrombin III binding sequence determined by hplc analysis in these latter fractions correlates well with the increases in anticoagulant activities observed upon fractionation.
  • Isolation and characterization of unidentified oligosaccharide products in these deamination populations showed the presence of a monosulfated (with D-glucuronic acid reducing terminal) and a disulfated trisaccharide {(alpha)-L-idopyranosyluronic acid-(1(--->)4)-2-acetamido-2-deoxy-(alpha)-D-glucopyranose-(1(--->)4)-D-glucuronic acid}, a trisulfated tetrasaccharide {(alpha)-L-idopyranosyluronic acid-(1(--->)4)-2-acetamido-2-deoxy-(alpha)-D-glucopyranose-(1(--->)4)-(beta)-D-glucopyranosyluronic acid-(1(--->)4)-2,5-anhydro-D-mannitol}, and oligosaccharides whose monosaccharide sequences were only partially characterized. These findings suggest that a far greater structural heterogeneity exists in the linear sequence of heparin monosaccharide units than currently recognized.
Graduation Semester
1981
Type of Resource
text
Permalink
http://hdl.handle.net/2142/67417

Owning Collections

Graduate Dissertations and Theses at Illinois PRIMARY
Graduate Theses and Dissertations at Illinois
Dissertations and Theses - Biochemistry