title: New insights on the specificity of heparin and heparan sulfate lyases from Flavobacterium heparinum revealed by the use of synthetic derivatives of K5 polysaccharide from E. coli and 2-O-desulfated heparin.
abstract: The capsular polysaccharide from E. Coli, strain K5 composed of ...-->4)beta-D-GlcA(1-->4)alpha-D-GlcNAc(1-->4)beta-D-GlcA (1-->..., chemically modified K5 polysaccharides, bearing sulfates at C-2 and C-6 of the hexosamine moiety and at the C-2 of the glucuronic acid residues as well as 2-O desulfated heparin were used as substrates to study the specificity of heparitinases I and II and heparinase from Flavobacterium heparinum. The natural K5 polysaccharide was susceptible only to heparitinase I forming deltaU-GlcNAc. N-deacetylated, N-sulfated K5 became susceptible to both heparitinases I and II producing deltaU-GlcNS. The K5 polysaccharides containing sulfate at the C-2 and C-6 positions of the hexosamine moiety and C-2 position of the glucuronic acid residues were susceptible only to heparitinase II producing deltaU-GlcNS,6S and deltaU,2S-GlcNS,6S respectively. These combined results led to the conclusion that the sulfate at C-6 position of the glucosamine is impeditive for the action of heparitinase I and that heparitinase II requires at least a C-2 or a C-6 sulfate in the glucosamine residues of the substrate for its activity. Iduronic acid-2-O-desulfated heparin was susceptible only to heparitinase II producing deltaU-GlcNS,6S. All the modified K5 polysaccharides as well as the desulfated heparin were not substrates for heparinase. This led to the conclusion that heparitinase II acts upon linkages containing non-sulfated iduronic acid residues and that heparinase requires C-2 sulfated iduronic acid residues for its activity.
authors.affiliation: Departamento de Bioquimica, Escoloa Paulista de Medicina, UNIFESP, São Paulo, S.P., Brazil. hbnader.bioq@unifesp.br
authors.forename: H B
authors.initials: HB
authors.lastname: Nader
authors.forename: E Y
authors.initials: EY
authors.lastname: Kobayashi
authors.forename: S F
authors.initials: SF
authors.lastname: Chavante
authors.forename: I L
authors.initials: IL
authors.lastname: Tersariol
authors.forename: R A
authors.initials: RA
authors.lastname: Castro
authors.forename: S K
authors.initials: SK
authors.lastname: Shinjo
authors.forename: A
authors.initials: A
authors.lastname: Naggi
authors.forename: G
authors.initials: G
authors.lastname: Torri
authors.forename: B
authors.initials: B
authors.lastname: Casu
authors.forename: C P
authors.initials: CP
authors.lastname: Dietrich
mesh_terms: D016667:Bacterial Capsules; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000737:chemistry; D005417:Flavobacterium / Q000737:chemistry*; D006493:Heparin / Q000737:chemistry*; D019763:Heparin Lyase / Q000737:chemistry*; D006497:Heparan Sulfate / Q000737:chemistry; D008969:Molecular Sequence Data; D011133:Polysaccharide-Lyases / Q000737:chemistry*; D011135:Polysaccharides, Bacterial / Q000737:chemistry*; D013379:Substrate Specificity
chemical_list: D006493:Heparin; D019763:Heparin Lyase; D006497:Heparan Sulfate; D011133:Polysaccharide-Lyases; D011135:Polysaccharides, Bacterial; C050673:capsular polysaccharide K5; C114663:heparinase II; C021349:heparitinsulfate lyase