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Identification of oligomeric domains within dermatan sulfate chains using differential enzymic treatments, derivatization with 2-aminoacridone and capillary electrophoresis.
Electrophoresis. 2001 Aug; 22(12):2458-63.E

Abstract

Galactosaminoglycans, i.e. dermatan sulfate (DS) and chondroitin sulfate, are linear heteropolysaccharides consisting of repeating disaccharide units of L-iduronic acid (L-IdoA) or D-glucuronic acid (D-GlcA) residues linked to N-acetyl-galactosamine. High-performance capillary electrophoresis (HPCE or CE) has been successfully used for determining the disaccharide composition of glycosaminoglycans. However, only limited information is available on how to identify oligomeric domains rich in D-GlcA or L-IdoA. The aim of this study was therefore to develop a rapid and accurate CE procedure by which such oligosaccharides can be determined together with the variously sulfated disaccharides. Isolated dermatan sulfates of human origin were separately digested with chondroitinases ABC, AC and B and the enzymic products were derivatized with 2-aminoacridone. CE analysis of these products was performed using a phosphate buffer, pH 3.0, and reversed polarity at 30 kV. The derivatization enabled their detection with laser-induced fluorescence (LIF) and UV at 260 nm at much higher sensitivity than the detection of nonderivatized delta-saccharides at 232 nm and therefore components undetectable at 232 nm were nicely detected after derivatization. Except for delta-disaccharides, altogether five distinct oligosaccharides with differences in charge density were identified. Depending on the lyase that produced these oligomers, information on the presence of L-IdoA- or D-GlcA-containing domains within the DS chain and the sulfation pattern of these oligomeric domains was obtained. This CE method could also be useful in studying the functional oligomeric domains in galactosaminoglycan chains.

Authors+Show Affiliations

Mitropoulou TN
Department of Chemistry, University of Patras, Greece.
Lamari F
No affiliation info available
Syrokou A
No affiliation info available
Hjerpe A
No affiliation info available
Karamanos NK
No affiliation info available

MeSH

AminoacridinesBacterial ProteinsChondroitin ABC LyaseChondroitin LyasesChondroitinases and Chondroitin LyasesDermatan SulfateDisaccharidesElectrophoresis, CapillaryFemaleGlucuronic AcidHumansIduronic AcidLeiomyomaMyometriumOligosaccharidesPolysaccharidesProtein Structure, TertiarySubstrate SpecificityUterine Neoplasms

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

11519950

Citation

Mitropoulou, T N., et al. "Identification of Oligomeric Domains Within Dermatan Sulfate Chains Using Differential Enzymic Treatments, Derivatization With 2-aminoacridone and Capillary Electrophoresis." Electrophoresis, vol. 22, no. 12, 2001, pp. 2458-63.
Mitropoulou TN, Lamari F, Syrokou A, et al. Identification of oligomeric domains within dermatan sulfate chains using differential enzymic treatments, derivatization with 2-aminoacridone and capillary electrophoresis. Electrophoresis. 2001;22(12):2458-63.
Mitropoulou, T. N., Lamari, F., Syrokou, A., Hjerpe, A., & Karamanos, N. K. (2001). Identification of oligomeric domains within dermatan sulfate chains using differential enzymic treatments, derivatization with 2-aminoacridone and capillary electrophoresis. Electrophoresis, 22(12), 2458-63.
Mitropoulou TN, et al. Identification of Oligomeric Domains Within Dermatan Sulfate Chains Using Differential Enzymic Treatments, Derivatization With 2-aminoacridone and Capillary Electrophoresis. Electrophoresis. 2001;22(12):2458-63. PubMed PMID: 11519950.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Identification of oligomeric domains within dermatan sulfate chains using differential enzymic treatments, derivatization with 2-aminoacridone and capillary electrophoresis. AU - Mitropoulou,T N, AU - Lamari,F, AU - Syrokou,A, AU - Hjerpe,A, AU - Karamanos,N K, PY - 2001/8/25/pubmed PY - 2002/2/8/medline PY - 2001/8/25/entrez SP - 2458 EP - 63 JF - Electrophoresis JO - Electrophoresis VL - 22 IS - 12 N2 - Galactosaminoglycans, i.e. dermatan sulfate (DS) and chondroitin sulfate, are linear heteropolysaccharides consisting of repeating disaccharide units of L-iduronic acid (L-IdoA) or D-glucuronic acid (D-GlcA) residues linked to N-acetyl-galactosamine. High-performance capillary electrophoresis (HPCE or CE) has been successfully used for determining the disaccharide composition of glycosaminoglycans. However, only limited information is available on how to identify oligomeric domains rich in D-GlcA or L-IdoA. The aim of this study was therefore to develop a rapid and accurate CE procedure by which such oligosaccharides can be determined together with the variously sulfated disaccharides. Isolated dermatan sulfates of human origin were separately digested with chondroitinases ABC, AC and B and the enzymic products were derivatized with 2-aminoacridone. CE analysis of these products was performed using a phosphate buffer, pH 3.0, and reversed polarity at 30 kV. The derivatization enabled their detection with laser-induced fluorescence (LIF) and UV at 260 nm at much higher sensitivity than the detection of nonderivatized delta-saccharides at 232 nm and therefore components undetectable at 232 nm were nicely detected after derivatization. Except for delta-disaccharides, altogether five distinct oligosaccharides with differences in charge density were identified. Depending on the lyase that produced these oligomers, information on the presence of L-IdoA- or D-GlcA-containing domains within the DS chain and the sulfation pattern of these oligomeric domains was obtained. This CE method could also be useful in studying the functional oligomeric domains in galactosaminoglycan chains. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/11519950/Identification_of_oligomeric_domains_within_dermatan_sulfate_chains_using_differential_enzymic_treatments_derivatization_with_2_aminoacridone_and_capillary_electrophoresis_ L2 - https://doi.org/10.1002/1522-2683(200107)22:12<2458::AID-ELPS2458>3.0.CO;2-8 DB - PRIME DP - Unbound Medicine ER -
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