Selective cleavage of heparin using aqueous 2-hydroxypyridine: production of an aldose-terminating fragment with high anticoagulant activity.
Biochem Biophys Res Commun
; 346(3): 946-57, 2006 Aug 04.
Article
em En
| MEDLINE
| ID: mdl-16781674
ABSTRACT
Unfractionated heparin (UFH) was partially depolymerized by heating at 115 degrees C with aqueous 2-hydroxypyridine. Compared to starting UFH, no significant loss of anticoagulant (anti-Xa) activity was observed. Products consisted of polysaccharide fragments and small quantities of ammonia, sulfate, and hexuronic acid. Fragments with aldose termini that reacted with [3H]NaBH4 (fragment A) were of relatively uniform size (6000 D) and increased as depolymerization time increased. Fragment A contained the anticoagulant activity, with 90-94% and 24-31% binding to Sepharose-thrombin and Sepharose-antithrombin, respectively. In contrast, a non-reducing fragment B that did not react with [3H]NaBH4 was more heterogeneous (6000-10,000 D) and did not have anticoagulant activity or Sepharose-antithrombin affinity. Given the polysaccharide 3H-incorporation, small release of monosaccharide products, and fragment A end-group analysis, thermolysis of UFH is likely limited to one site per molecule when protected by 2-hydroxypyridine. Thus, an anticoagulant fragment A is hydrolytically released from UFH leaving a variable-length fragment B complete with linkage region.
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Base de dados:
MEDLINE
Assunto principal:
Polissacarídeos
/
Piridonas
/
Heparina
/
Anticoagulantes
Tipo de estudo:
Prognostic_studies
Limite:
Animals
Idioma:
En
Ano de publicação:
2006
Tipo de documento:
Article