Subscribe to RSS
DOI: 10.1055/s-0038-1649706
Interaction of Immobilised Unfractionated and LMW Heparins with Proteins in Whole Human Plasma
Publication History
Received 17 February 1993
Accepted after revision 12 July 1993
Publication Date:
06 July 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/199306/lookinside/thumbnails/10-1055-s-0038-1649706-1.jpg)
Summary
The profile of proteins bound to immobilised heparins in hirudin-anticoagulated human plasma was analysed. In molar terms, antithrombin III was the most abundant protein bound to therapeutic doses of unfractionated heparin (Mr = 12,000), whereas heparin cofactor II constituted <1% of the protein bound. Histidine-rich glycoprotein was the only plasma protein likely to influence anticoagulant activity by direct competition with antithrombin III, though significant quantities of complement Factor H, fibrinogen, fibronectin, vitronectin and apolipo-protein B were also detected. Only traces of von Willebrand factor, complement factor I, inter-α-trypsin inhibitor, a2-macro-globulin, serum amyloid P and transferrin were identified, and neither thrombospondin nor platelet factor 4 were measurable. Binding of both antithrombin III and histidine-rich glycoprotein varied with the ratio of heparin to plasma. Clexane (Mr = 4,500) also bound anti thrombin III, but both histidine-rich glycoprotein and vitronectin were quantitatively significant neutralising proteins. Neutralising proteins dominated the binding profile for Oligo H (Mr = 2,200).
-
References
- 1 Choay J, Petitou M, Lormeau JC, Sinay P, Casu B, Gatti G. Structure activity relationship in heparin: a synthetic pentasaccharide with high affinity for antithrombin III and eliciting high anti-factor Xa activity. Biochem Biophys Res Comm 1983; 116: 492-499
- 2 Tollefsen DM, Blank MK. Detection of a new heparin-dependent inhibitor of thrombin in human plasma. J Clin Invest 1981; 68: 589-596
- 3 Lane DA, Denton J, Flynn AM, Thunberg L, Lindahl U. Anticoagulant activities of heparin oligosaccharides and their neutralization by platelet factor 4. Biochem J 1984; 218: 725-732
- 4 Bray B, Lane DA, Freyssinet JM, Pejler G, Lindahl U. Antithrombin activities of heparin. Effect of saccharide chain length on thrombin inhibition by heparin cofactor II and by antithrombin. Biochem J 1989; 262: 225-232
- 5 Andersson L-O, Barrowcliffe TW, Holmer E, Johnson EA, Soder-strom G. Molecular weight dependency of the heparin potentiated inhibition of thrombin and activated factor X. Effect of heparin neutralization in plasma. Thromb Res 1979; 15: 531-541
- 6 McKay EJ, Laurell C-B. The interaction of heparin with plasma proteins. Demonstration of different binding sites for antithrombin III complexes and antithrombin III. J Lab Clin Med 1980; 95: 69-80
- 7 Young E, Prins M, Levine MN, Hirsh J. Heparin binding to plasma proteins, an important mechanism for heparin resistance. Thromb Haemostas 1992; 67: 639-643
- 8 Barrowcliffe TW, Le Shirley Y. The effect of calcium chloride on anti-Xa activity of heparin and its molecular weight fractions. Thromb Haemostas 1989; 62: 950-954
- 9 Lijnen HR, Hoylaerts M, Collen D. Heparin-binding properties of human histidine-rich glycoprotein. Mechanism and role in the neutralization of heparin in plasma. J Biol Chem 1983; 258: 3803-3808
- 10 Sim E, Palmer MS, Puklavec M, Sim RB. Monoclonal antibodies against the complement control protein Factor H (β1H). Biosci Rep 1983; 3: 1119-1131
- 11 Gray E, Watton J, Cesmeli S, Barrowcliffe TW, Thomas DP. Experimental studies on a recombinant hirudin, CGP 39393. Thromb Haemostas 1991; 65: 355-359
- 12 Dawes J. Measurement of the affinities of heparins, naturally occurring glycosaminoglycans, and other sulfated polysaccharides for antithrombin III and thrombin. Anal Biochem 1988; 174: 177-186
- 13 Hunter WM. Simplified solid-phase radioimmunoassay without centrifugation. Acta Endocrinol 1977; 85 (Suppl. 212) 463
- 14 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4 . Nature 1970; 227: 680-685
- 15 Kazama Y, Koide T. Modulation of protein C inhibitor activity by histidine-rich glycoprotein and platelet factor 4: Role of zinc and calcium ions in the heparin-neutralising ability of histidine-rich glycoprotein. Thromb Haemostas 1992; 67: 50-55
- 16 Peterson CB, Morgan WT, Blackburn MN. Histidine-rich glycoprotein modulation of the anticoagulant activity of heparin. Evidence for a mechanism involving competition with both antithrombin and thrombin for heparin binding. J Biol Chem 1987; 262: 7567-7574
- 17 Griffith MJ. Heparin catalysed inhibitor/protease reactions: kinetic evidence for a common mechanism of action of heparin. Proc Natl Acad Sci USA 1983; 80: 5460-5464
- 18 Olson ST, Srinavasan KR, Bjork I, Shore JD. Binding of high affinity heparin to antithrombin III. Stopped flow kinetic studies of the binding interaction. J Biol Chem 1981; 256: 11073-11079
- 19 Yamada KM, Kennedy DW, Kimata K, Pratt RM. Characterization of fibronectin interactions with glycosaminoglycans and identification of active proteolytic fragments. J Biol Chem 1980; 255: 6055-6053
- 20 Preissner KT, Muller-Berghaus G. Neutralization and binding of heparin by S-protein/vitronectin in the inhibition of factor Xa by antithrombin III. Involvement of an inducible heparin-binding domain of S-protein/vitronectin. J Biol Chem 1987; 262: 12247-12253
- 21 Engesser L, Kluft C, Briët E, Brommer EJP. Familial elevation of plasma histidine-rich glycoprotein in a family with thrombophilia. Br J Haemat 1987; 67: 355-358
- 22 Preissner KT, Zwicker L, Muller-Berghaus G. Formation, characterization and detection of a ternary complex between S protein, thrombin and antithrombin III in serum. Biochem J 1987; 243: 105-111
- 23 Preissner KT, Sie P. Modulation of heparin cofactor II function by S-protein (vitronectin) and formation of a ternary S protein-thrombinheparin cofactor II complex. Thromb Haemostas 1988; 60: 399-406
- 24 Plow EF, Ginsberg MH. Cellular adhesion: GPIIb-IIIa as a prototypic adhesion receptor. Prog Hemost Thromb 1989; 9: 117-156
- 25 Nievelstein PF, de Groot PG, D’Alessio P, Heijnen HF, Orlando E, Sixma JJ. Platelet adhesion to vascular cells. The role of exogenous von Willebrand factor in platelet adhesion. Arteriosclerosis 1990; 10: 462-469
- 26 Podack ER, Muller-Eberhard HJ. Isolation of human S-protein, an inhibitor of the membrane attack complex of complement. J Biol Chem 1979; 254: 9808-9814
- 27 Hamilton KK, Hattori R, Esmon CT, Sims PJ. Complement proteins C5b-9 induce vesiculation of the endothelial plasma membrane and expose a catalytic surface for assembly of the prothrombinase enzyme complex. J Biol Chem 1990; 265: 3809-3814
- 28 Meri S, Pangburn MK. Discrimination between activators and nonactivators of the alternative pathway of complement: regulation via a sialic acid/poly anion binding site on factor H. Proc Natl Acad Sci USA 1990; 87: 3982-3986
- 29 Dawes J, Prowse CV, Pepper DS. Absorption of heparin, LMW heparin and SP54 after subcutaneous injection, assessed by competitive binding assay. Thromb Res 1986; 44: 683-693
- 30 Linhardt RJ, Rice KG, Merchant ZM, Kim YS, Lohse DL. Structure and activity of a unique heparin-derived hexasaccharide. J Biol Chem 1986; 261: 14448-14454
- 31 Lane DA, Pejler G, Flynn AM, Thompson EA, Lindahl U. Neutralization of heparin-related saccharides by histidine-rich glycoprotein and platelet factor 4. J Biol Chem 1986; 261: 3980-3986
- 32 Lane DA, Flynn AM, Pejler G, Lindahl U, Choay J, Preissner K. Structural requirements for the neutralisation of heparin-like saccharides by complement S protein/vitronectin. J Biol Chem 1987; 262: 16343-16348
- 33 Ogamo A, Nagai A, Nagasawa K. Binding of heparin fractions and other polysulfated polysaccharides to plasma fibronectin: effects of molecular size and degree of sulfation of polysaccharides. Biochim Biophys Acta 1985; 841: 30-41