Examination of the substrate specificity of heparin and heparan sulfate lyases.
Affiliation
Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242.Issue Date
1990-03-13
Metadata
Show full item recordAbstract
We have examined the activities of different preparations of heparin and heparan sulfate lyases from Flavobacterium heparinum. The enzymes were incubated with oligosaccharides of known size and sequence and with complex polysaccharide substrates, and the resulting degradation products were analyzed by strong-anion-exchange high-performance liquid chromatography and by oligosaccharide mapping using gradient polyacrylamide gel electrophoresis. Heparinase (EC 4.2.2.7) purified in our laboratory and a so-called Heparinase I (Hep I) from a commercial source yielded similar oligosaccharide maps with heparin substrates and displayed specificity for di- or trisulfated disaccharides of the structure----4)-alpha-D-GlcNp2S(6R)(1----4)-alpha-L-IdoAp2S( 1----(where R = O-sulfo or OH). Oligosaccharide mapping with two different commercial preparations of heparan sulfate lyase [heparitinase (EC 4.2.2.8)] indicated close similarities in their depolymerization of heparan sulfate. Furthermore, these enzymes only degraded defined oligosaccharides at hexosaminidic linkages with glucuronic acid:----4)-alpha-D-GlcNpR(1----4)-beta-D-GlcAp(1----(where R = N-acetamido or N-sulfo). The enzymes showed activity against solitary glucuronate-containing disaccharides in otherwise highly sulfated domains including the saccharide sequence that contains the antithrombin binding region in heparin. A different commercial enzyme, Heparinase II (Hep II), displayed a broad spectrum of activity against polysaccharide and oligosaccharide substrates, but mapping data indicated that it was a separate enzyme rather than a mixture of heparinase and heparitinase/Hep III. When used in conjunction with the described separation procedures, these enzymes are powerful reagents for the structural/sequence analysis of heparin and heparan sulfate.Citation
Examination of the substrate specificity of heparin and heparan sulfate lyases. 1990, 29 (10):2611-7 BiochemistryJournal
BiochemistryDOI
10.1021/bi00462a026PubMed ID
2334685Type
ArticleLanguage
enISSN
0006-2960ae974a485f413a2113503eed53cd6c53
10.1021/bi00462a026
Scopus Count
Collections
Related articles
- Specificity studies on the heparin lyases from Flavobacterium heparinum.
- Authors: Desai UR, Wang HM, Linhardt RJ
- Issue date: 1993 Aug 17
- Purification and substrate specificity of heparitinase I and heparitinase II from Flavobacterium heparinum. Analyses of the heparin and heparan sulfate degradation products by 13C NMR spectroscopy.
- Authors: Nader HB, Porcionatto MA, Tersariol IL, Pinhal MA, Oliveira FW, Moraes CT, Dietrich CP
- Issue date: 1990 Oct 5
- Isolation of the porcine heparin tetrasaccharides with glucuronate 2-O-sulfate. Heparinase cleaves glucuronate 2-O-sulfate-containing disaccharides in highly sulfated blocks in heparin.
- Authors: Yamada S, Murakami T, Tsuda H, Yoshida K, Sugahara K
- Issue date: 1995 Apr 14
- Purification and characterization of heparin lyases from Flavobacterium heparinum.
- Authors: Lohse DL, Linhardt RJ
- Issue date: 1992 Dec 5
- Substrate specificity of the heparin lyases from Flavobacterium heparinum.
- Authors: Desai UR, Wang HM, Linhardt RJ
- Issue date: 1993 Nov 1