Heparin dodecasaccharide containing two antithrombin-binding pentasaccharides: structural features and biological properties

• Published in: The Journal of biological chemistry Vol. 288; no. 36; pp. 25895 - 25907
• Main Authors: Viskov, Christian, Elli, Stefano, Urso, Elena, Gaudesi, Davide, Mourier, Pierre, Herman, Frederic, Boudier, Christian, Casu, Benito, Torri, Giangiacomo, Guerrini, Marco
• Format: Journal Article
• Language: English
• Published: United States American Society for Biochemistry and Molecular Biology 06.09.2013
• Subjects: Carbohydrate Sequence; Factor Xa - chemistry; Fibrinolytic Agents - chemical synthesis; Fibrinolytic Agents - chemistry; Glycobiology and Extracellular Matrices; Heparin; Humans; Life Sciences; Magnetic Resonance Spectroscopy; Oligosaccharides - chemical synthesis; Oligosaccharides - chemistry; Mass Spectrometry (MS); Glycosaminoglycan; NMR; Heparin; Antithrombin; 3-O-Sulfation
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M113.485268

The antithrombin (AT) binding properties of heparin and low molecular weight heparins are strongly associated to the presence of the pentasaccharide sequence AGA*IA (A(NAc,6S)-GlcUA-A(NS,3,6S)-I(2S)-A(NS,6S)). By using the highly chemoselective depolymerization to prepare new ultra low molecular weight heparin and coupling it with the original separation techniques, it was possible to isolate a polysaccharide with a biosynthetically unexpected structure and excellent antithrombotic properties. It consisted of a dodecasaccharide containing an unsaturated uronate unit at the nonreducing end and two contiguous AT-binding sequences separated by a nonsulfated iduronate residue. This novel oligosaccharide was characterized by NMR spectroscopy, and its binding with AT was determined by fluorescence titration, NMR, and LC-MS. The dodecasaccharide displayed a significantly increased anti-FXa activity compared with those of the pentasaccharide, fondaparinux, and low molecular weight heparin enoxaparin.