Structure of the antithrombin-binding site in heparin.

Lindahl, Ulf; Bäckström, G.; Höök, Magnus; Thunberg, Lennart; Fransson, Lars‐Âke; Linker, Alfred

doi:10.1073/pnas.76.7.3198

Cited by 321 publications

(174 citation statements)

References 16 publications

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“…Several of the biological activities of proteoglycans are attributable to the GAGs. For example, heparin binds anti-thrombin III (1), and heparin and HS bind fibroblast growth factor (2) and various chemokines (3)(4)(5). Furthermore, CS binds a subset of chemokines, such as RANTES (regulated on activation normal T cell expressed and secreted) (6) and PF-4 (platelet factor-4) (7).…”

mentioning

confidence: 99%

“…They provide structural integrity to tissues and mediate cell proliferation, differentiation, and migration. Proteoglycans consist of a core protein to which one or more glycosaminoglycan (GAG) 1 side chains are covalently attached. There are several types of GAGs, which include heparin or heparan sulfate (HS), chondroitin sulfate (CS), dermatan sulfate (CS B), and keratan sulfate (KS).…”

mentioning

confidence: 99%

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Binding of a Large Chondroitin Sulfate/Dermatan Sulfate Proteoglycan, Versican, to L-selectin, P-selectin, and CD44

Kawashima¹,

Hirose²,

Hirose³

et al. 2000

Journal of Biological Chemistry

242

194

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Here we show that a large chondroitin sulfate proteoglycan, versican, derived from a renal adenocarcinoma cell line ACHN, binds L-selectin, P-selectin, and CD44. The binding was mediated by the interaction of the chondroitin sulfate (CS) chain of versican with the carbohydrate-binding domain of L-and P-selectin and CD44. The binding of versican to L-and P-selectin was inhibited by CS B, CS E, and heparan sulfate (HS) but not by any other glycosaminoglycans tested. On the other hand, the binding to CD44 was inhibited by hyaluronic acid, chondroitin (CH), CS A, CS B, CS C, CS D, and CS E but not by HS or keratan sulfate. A crossblocking study indicated that L-and P-selectin recognize close or overlapping sites on versican, whereas CD44 recognizes separate sites. We also show that soluble L-and P-selectin directly bind to immobilized CS B, CS E, and HS and that soluble CD44 directly binds to immobilized hyaluronic acid, CH, and all the CS chains examined. Consistent with these results, structural analysis showed that versican is modified with at least CS B and CS C. Thus, proteoglycans sufficiently modified with the appropriate glycosaminoglycans should be able to bind L-selectin, P-selectin, and/or CD44.

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mentioning

confidence: 99%

mentioning

confidence: 99%

Binding of a Large Chondroitin Sulfate/Dermatan Sulfate Proteoglycan, Versican, to L-selectin, P-selectin, and CD44

Kawashima¹,

Hirose²,

Hirose³

et al. 2000

Journal of Biological Chemistry

242

194

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“…Thus, in heparan sulphate 5 consecutive repeats of this kind may be present to a considerable extent. This is likely to be a basis for the significant anticoagulant activity displayed by this material [33].…”

Section: Discussionmentioning

confidence: 99%

Structural Studies on Heparan Sulphates

Fransson¹,

Sjöberg²,

Havsmark³

1980

European Journal of Biochemistry

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Three heparan sulphate fractions were subjected to degradation by periodate oxidation and alkaline elimination. The starting materials were one low-sulphated fraction rich in glucuronic acid and iV-acetylglucosamine (I), one fraction (IV) that was medium-suphated, contained similar proportions of iV-acetyl and iV-sulphate as well as of glucuronic and iduronic acid and one fraction (V) that was oversulphated, contained an excess of iV-sulphate and was rich in iduronic acid. Selective periodate oxidation of glucuronic acid residues within iV-acetylated regions followed by scission in alkali yielded three categories of fragments that were isolated by gel and ion-exchange chromatography. (a) iV-Acetylglucosamine-R compounds where R is the remnant of an oxidised and degraded glucuronic acid residue. (b) Medium-sized oligosaccharides, of the general structure glucosamine-(glycuronic acid-glucosamine),-R where n = 1 -4, contained largely glucuronic acid associated with glucosamines that were iV-sulphated, iV-acetylated or unsubstituted. In saccharides where n = 4 indications of an alternating arrangement of iV-sulphate and iV-acetyl groups as well as of iduronic and glucuronic acid residues were obtained. The glucuronic acid residues of the oligosaccharide fragments were susceptible to reoxidation with periodate. The smaller saccharides were not depolymerised by HNO, despite the presence of iV-sulphate groups. (c) Longer oligosaccharide fragments (n 2 5) that were highly sulphated and contained heparin-like repeating units, i. e. iduronic acid -glucosamine-iV-sulphate with ester-sulphate on both sugars. These oligosaccharides were susceptible to deaminative cleavage.

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“…However, the existence of a small number of high-molecular-weight heparin molecules with even somewhat higher binding constants is suggested by the shape of the affinitychromatographic elution patterns. The observed range of affinities may be due to a variation of the structure within the antithrombin-binding site of the high-affinity heparin molecules (Lindahl et al, 1979). However, a variation of the number of negatively charged sulphate groups outside the 1981 binding area of the molecules could also result in different affinities for antithrombin owing to a general electrostatic effect.…”

Section: Molecular Weightsmentioning

confidence: 99%

Binding to antithrombin of heparin fractions with different molecular weights

Danielsson

Björk

1981

Biochemical Journal

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The interaction between bovine antithrombin, a plasma proteinase inhibitor, and heparin species of different molecular weights was studied. A commercial heparin preparation was divided by gel chromatography into a number of fractions with average molecular weights ranging from 6000 to 34 700. Each of these fractions was further fractionated by affinity chromatography on matrix-bound antithrombin. In the latter procedure, those heparin fractions that had molecular weights lower than about 14000 were separated into three peaks. The material in the first of these was not adsorbed on the column, and the other two peaks corresponded to the low-affinity and high-affinity peaks described previously. In contrast, high-molecular-weight heparin samples gave only the low-affinity and high-affinity fractions. U.v. difference absorption studies showed that the non-adsorbed heparin fraction bound to antithrombin in solution with a binding constant at physiological ionic strength only slightly lower than that of low-affinity heparin. The division between the two fractions thus is arbitrary and only dependent on the conditions selected for the affinity-chromatography experiment. Stoicheiometries and binding constants for the binding of several high-affinity heparin species to antithrombin were determined by fluorescence titrations. High-affinity heparin fractions of equal elution positions in the beginning of the peaks of the affinity chromatographies, but with different molecular weights, showed stoicheiometries that were not experimentally distinguishable from 1:1 and also had no appreciable differences in binding constants. However, the anticoagulant activities, calculated on a molar basis, of these fractions increased markedly with molecular weight, a behaviour that thus cannot be explained by differences in the binding of the fractions to antithrombin. In contrast, high-affinity samples of similar molecular weights, which were eluted at increasing ionic strengths from matrix-linked antithrombin, were found to have an increasing proportion of chains with two binding sites for antithrombin and also to have progressively higher binding constants. These binding properties at least partly explain the increasing anticoagulant activities that were observed for these fractions.

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Structure of the antithrombin-binding site in heparin.

Cited by 321 publications

References 16 publications

Binding of a Large Chondroitin Sulfate/Dermatan Sulfate Proteoglycan, Versican, to L-selectin, P-selectin, and CD44

Binding of a Large Chondroitin Sulfate/Dermatan Sulfate Proteoglycan, Versican, to L-selectin, P-selectin, and CD44

Structural Studies on Heparan Sulphates

Binding to antithrombin of heparin fractions with different molecular weights

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