High-performance liquid chromatographic/mass spectrometric studies on the susceptibility of heparin species to cleavage by heparanase.

摘要: Heparanase is an endo-β -D-glucuronidase that cleaves the heparan sulfate chains of proteoglycans and implicated in angiogenesis metastasis. With aim establishing a simple reliable method for studying susceptibility heparin/ oligosaccharides to be cleaved by heparanase, on-line ion pair reversed-phase high-performance liquid chromatographic/electrospray ionization mass spectrometric was set up. The works micromolar range concentration does not require derivatization substrate or products. It based on identification ofoligosaccharide fragments generated heparanase their quantification with reference internal heparin disaccharide standard. Substrates were (1) synthetic pentasaccharides GlcN NS,6S - GlcA NS,3S,6S IdoA 2S OMe (AGA*IA M ) (AGAIA ), corresponding heparin/heparan active site antithrombin, same sequence devoid 3-O-sulfate group central glucosamine, respectively; (2) two natural octasaccharides containing AGA*IA different locations along chain. exhibited higher cleavage respect octasaccharides. commercial availability makes it ideal determine specific activity preparations. present could also used rapid screening potential inhibitors.