title: Determining the extent of heparan sulfate depolymerisation following heparin lyase treatment.
abstract: The depolymerisation of porcine mucosal heparan sulfate under the action of heparin lyases and analysis by size-exclusion chromatography (SEC) is described. Heparan sulfate treated to enzymic bond scission producing a Δ4,5 double-bond and quantified by SEC with ultraviolet-visible (UV) spectroscopic detection (230nm) indicated that the majority of the biopolymer (>85%) was reduced to disaccharides (degree of polymerisation (DP)=2). However, analysis of the SEC eluant using refractive index (RI), which reflects the mass contribution of the oligosaccharides rather than the molar response of a UV chromophore, indicated that a considerable proportion of the digested HS, up to 43%, was present with DP >2. This was supported by a mass balance analysis. These results contradict the accepted literature where "complete digestion" is routinely reported. Herein we report on the composition and methodology utilised to ascertain the extent of depolymerization and disaccharide composition of this important biopolymer.
authors: The Ferrier Research Institute, Victoria University of Wellington, 69 Gracefield Road, Lower Hutt, 5040, New Zealand.
authors: Susan M
authors: SM
authors: Carnachan
authors: The Ferrier Research Institute, Victoria University of Wellington, 69 Gracefield Road, Lower Hutt, 5040, New Zealand.
authors: Tracey J
authors: TJ
authors: Bell
authors: The Ferrier Research Institute, Victoria University of Wellington, 69 Gracefield Road, Lower Hutt, 5040, New Zealand.
authors: Ian M
authors: IM
authors: Sims
authors: Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), 8A Biomedical Grove, #06-06 Immunos, Singapore 138648, Singapore, Singapore.
authors: Raymond A A
authors: RAA
authors: Smith
authors: Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), 8A Biomedical Grove, #06-06 Immunos, Singapore 138648, Singapore, Singapore.
authors: Victor
authors: V
authors: Nurcombe
authors: Institute of Medical Biology, Agency for Science, Technology and Research (A*STAR), 8A Biomedical Grove, #06-06 Immunos, Singapore 138648, Singapore, Singapore.
authors: Simon M
authors: SM
authors: Cool
authors: The Ferrier Research Institute, Victoria University of Wellington, 69 Gracefield Road, Lower Hutt, 5040, New Zealand. Electronic address: Simon.Hinkley@vuw.ac.nz.
authors: Simon F R
authors: SFR
authors: Hinkley
mesh_terms: D000818:Animals; D002417:Cattle; D019763:Heparin Lyase / Q000737:chemistry*; D006497:Heparan Sulfate / Q000737:chemistry*; D012754:Sharks; D013552:Swine
chemical_list: D019763:Heparin Lyase; D006497:Heparan Sulfate
keywords: Chromatographic analysis; Glucoasaminoglycan; Heparan sulfate; Heparin lyase