title: Differences in the nonreducing ends of heparan sulfates excreted by patients with mucopolysaccharidoses revealed by bacterial heparitinases: a new tool for structural studies and differential diagnosis of Sanfilippo's and Hunter's syndromes.
abstract: Enzymatic and chemical analyses of the structures of heparan sulfates excreted in the urine by patients with Sanfilippo's and Hunter's syndromes revealed that their nonreducing ends differ from each other and reflect the enzyme deficiency of the syndromes. The heparan sulfates from the different syndromes were treated with heparitinase II, crude enzyme extracts from Flavobacterium heparinum, and nitrous acid degradation. The heparan sulfates from patients with Sanfilippo A (deficient in heparan N-sulfatase) and Sanfilippo B (deficient in alpha-N-acetylglucosaminidase) were degraded with heparitinase II producing, besides unsaturated disaccharides, substantial amounts of glucosamine N-sulfate and N-acetylglucosamine, respectively. The heparan sulfate from patients with Hunter's syndrome (deficient in iduronate sulfatase) were degraded by heparitinase II or crude enzyme extracts to several products, including two saturated disaccharides containing a sulfated uronic acid at their nonreducing ends. The heparan sulfate from patients with Sanfilippo's C syndrome (deficient in acetyl Co-A: alpha-glucosaminide acetyltransferase) produced, by action of heparitinase II, among other products, two sulfated trisaccharides containing glucosamine with a nonsubstituted amino group. In addition to providing a new tool for the differential diagnosis of the mucopolysaccharidoses, these results bring new insights into the specificity of the heparitinases from Flavobacterium heparinum.
authors.affiliation: Department of Biochemistry, Escola Paulista de Medicina, Universidade Federal de São Paulo, Brazil.
authors.forename: L
authors.initials: L
authors.lastname: Toma
authors.forename: C P
authors.initials: CP
authors.lastname: Dietrich
authors.forename: H B
authors.initials: HB
authors.lastname: Nader
mesh_terms: D015415:Biomarkers / Q000652:urine; D002240:Carbohydrate Sequence; D002809:Chondroitin Sulfates / Q000652:urine*; D003871:Dermatan Sulfate / Q000652:urine*; D003937:Diagnosis, Differential; D004587:Electrophoresis, Agar Gel; D006497:Heparan Sulfate / Q000737:chemistry / Q000652:urine*; D006801:Humans; D008969:Molecular Sequence Data; D016532:Mucopolysaccharidosis II / Q000175:diagnosis* / Q000652:urine; D009084:Mucopolysaccharidosis III / Q000175:diagnosis* / Q000652:urine; D011133:Polysaccharide-Lyases / Q000378:metabolism*; D012016:Reference Values; D013379:Substrate Specificity
chemical_list: D015415:Biomarkers; D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D006497:Heparan Sulfate; D011133:Polysaccharide-Lyases; C021349:heparitinsulfate lyase