Biosynthesis of heparin. Different molecular forms of O-sulfotransferases.

Wlad, Hanna; Maccarana, Marco; Eriksson, Inger; Kjellén, Lena; Lindahl, Ulf

doi:10.1016/s0021-9258(17)31423-0

Cited by 34 publications

(5 citation statements)

References 23 publications

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“…The supernatant was discarded, and the pellet was dissolved in 100 µL of water. The 35 S-labeled polysaccharide was recovered after centrifugation through Sephadex G-25 as described previously (46). The fluid recovered in the centrifuge tubes contained the 35 Slabeled polysaccharide which was quantified by liquid scintillation counting.…”

Section: Methodsmentioning

confidence: 99%

Substrate Specificity of the Heparan Sulfate Hexuronic Acid 2-O-Sulfotransferase

et al. 2001

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The interaction of heparan sulfate with different ligand proteins depends on the precise location of O-sulfate groups in the polysaccharide chain. We have previously shown that overexpression in human kidney 293 cells of a mouse mastocytoma 2-O-sulfotransferase (2-OST), previously thought to catalyze the transfer of sulfate from 3'-phosphoadenosine 5'-phosphosulfate to C2 of L-iduronyl residues, preferentially increases the level of 2-O-sulfation of D-glucuronyl units [Rong, J., Habuchi, H., Kimata, K., Lindahl, U., and Kusche-Gullberg, M. (2000) Biochem. J. 346, 463-468]. In the study presented here, we further investigated the substrate specificity of the mouse mastocytoma 2-OST. Different polysaccharide acceptor substrates were incubated with cell extracts from 2-OST-transfected 293 cells together with the sulfate donor 3'-phosphoadenosine 5'-phospho[(35)S]sulfate. Incubations with O-desulfated heparin, predominantly composed of [(4)alphaIdoA(1)-(4)alphaGlcNSO(3)(1)-](n)(), resulted in 2-O-sulfation of iduronic acid. When, on the other hand, an N-sulfated capsular polysaccharide from Escherichia coli K5, with the structure [(4)betaGlcA(1)-(4)alphaGlcNSO(3)(1)-](n)(), was used as an acceptor, sulfate was transferred almost exclusively to C2 of glucuronic acid. Substrates containing both iduronic and glucuronic acid residues in about equal proportions strongly favored sulfation of iduronic acid. In agreement with these results, the 2-OST was found to have a approximately 5-fold higher affinity for iduronic acid-containing substrate disaccharide units (K(m) approximately 3.7 microM) than for glucuronic acid-containing substrate disaccharide units (K(m) approximately 19.3 microM).

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Section: Methodsmentioning

confidence: 99%

Substrate Specificity of the Heparan Sulfate Hexuronic Acid 2-O-Sulfotransferase

et al. 2001

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“…6 These were assigned tentatively as: IdoA2OH-GlcNS,3S,6S-IdoA2S and GlcA-GlcNS,3S,6S-IdoA2S for the synthesis of glucosamine 2,3,6 tri-sulfate residues and IdoA-GlcNS,3S-IdoA, IdoA2S-GlcNS,3S-IdoA2S and GlcA-GlcNS,3S-IdoA2S in the case of GlcNS,3S, based on their relative abundance in heparin. 39 The disaccharide substrates for 3-O-sulfation can therefore be represented as; 011, G11, 010, 110 and G10, which also all lie on the same major branch of the proposed biosynthetic scheme, originating from G10 [Fig. 2].…”

Section: The Common Substrates For the 3-o-sulfation Of Glucosamine A...mentioning

confidence: 99%

A highly efficient tree structure for the biosynthesis of heparan sulfate accounts for the commonly observed disaccharides and suggests a mechanism for domain synthesis

Rudd¹,

Yates²

2012

Mol. BioSyst.

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The form of the biosynthetic pathway of the biologically and medically important polysaccharides heparan sulfate (HS) and the closely related heparin remain obscure despite significant progress characterising the biosynthetic machinery. Considering possible biosynthetic schemes using a graph approach and applying known constraints of enzyme order and specificity, a previously unreported system with a highly efficient tree structure emerged with two features: (1) All commonly occurring HS disaccharides could be synthesised through a common route, the major branch. (2) The least common disaccharides also occurred on a separate common branch, termed here the minor branch. This suggested that the relative abundance of these two sets of structures were the result of the specificity of a single enzyme (HS epimerase) acting at an early point in the scheme, to convert GlcA-GlcNS to IdoA-GlcNS in preference to converting GlcA-GlcNAc to IdoA-GlcNAc. A third key finding was that the common substrates for 3-O-sulfation all lie on the same (major) branch. The proposed scheme is consistent with a wide body of experiments comprising both biochemical data and results from HS biosynthetic enzyme knockout experiments in the literature. The major branch also contains a bifurcation, providing a choice of two distinct backbone geometries with the same charge. Further development of this novel biosynthetic scheme, in which frame shifts in the site of action of the enzymes were permitted to occur, while maintaining their order of action, suggested a mechanism by which domains could be generated, or further modification blocked. The relationship between the proposed pathway and the geometric and charge possibilities it allows were also explored.

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“…E-2-2. Heparan Sulfate 2-O-and 6-O-Sulfotransferase (HS2ST and HS6ST) An O-sulfotransferase, which transfers sulfate to N/Odesulfated and N-resulfated heparin (CDSNS-heparin), was purified from heparin-producing mouse mastocytoma (60). The purified enzyme showed both N-sulfoglucosaminyl 6-Osulfotransferase and iduronyl 2-O-sulfotransferase activities.…”

Section: E Modification Of Polysaccharide Enzymes Responsible For the Formation Of Diverged Structuresmentioning

confidence: 99%

Biosynthesis of Heparan Sulfate and Heparin. How Are the Multifunctional Glycosaminoglycans Built up?

Habuchi¹,

Habuchi²,

Kimata³

1998

Trends in Glycoscience and Glycotechnology

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Biosynthesis of heparin. Different molecular forms of O-sulfotransferases.

Cited by 34 publications

References 23 publications

Substrate Specificity of the Heparan Sulfate Hexuronic Acid 2-O-Sulfotransferase

Substrate Specificity of the Heparan Sulfate Hexuronic Acid 2-O-Sulfotransferase

A highly efficient tree structure for the biosynthesis of heparan sulfate accounts for the commonly observed disaccharides and suggests a mechanism for domain synthesis

Biosynthesis of Heparan Sulfate and Heparin. How Are the Multifunctional Glycosaminoglycans Built up?

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