Antithrombin and first complement protein recognize the same active heparin fraction.
Antithrombin (AT) high affinity of unfractionated heparin (UFH) resides in a specific pentasaccharide sequence. Heparin also regulates complement activity on the classical and the alternative pathways. Most experimental pieces of evidence accumulated show that these important activities reside in different segments of the heparin molecule. We demonstrated in previous papers that a low ionic strength and the presence of calcium ions are essential to detect specific interactions between glycosaminoglycans and proteins. Then these very strict conditions were used, and we demonstrated that the first protein complex of the human complement cascade recognizes in the UFH a fraction with very high anticoagulant activity. After isolation from the precipitate of the interaction, this fraction of heparin also contained the pentasaccharide sequence responsible for the great affinity with AT: in fact, it was strongly bound to a resin of AT agarose, and to detach it, an ionic strength of 0.6 M sodium chloride was required. In this way, the heparin regions responsible for the anticoagulant activity and also for the effects over the complement system were identified on the same short segment of the heparin molecule, which includes the active fraction of the glycosaminoglycan. The differences with early results could be explained by our experimental conditions of low ionic strength and the presence of calcium ions used for the interaction of the protein and the glycosaminoglycan.
Calabrese
Graciela C
GC
Departamento de Ciencias Biológicas, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 954/956 Buenos Aires 1110, Argentina. gcalebe@ffyb.uba.ar
Recondo
Eduardo F
EF
Fernandez de Recondo
Marta E
ME
D000925:Anticoagulants / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D000990:Antithrombin III / Q000378:metabolism*; D001665:Binding Sites; D002118:Calcium; D011232:Chemical Precipitation; D002846:Chromatography, Affinity; D003172:Complement C1 / Q000378:metabolism*; D006493:Heparin / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D009994:Osmolar Concentration; D011485:Protein Binding
D000925:Anticoagulants; D003172:Complement C1; D000990:Antithrombin III; D006493:Heparin; D002118:Calcium