Identification of an extended <i>N</i>-acetylated sequence adjacent to the protein-linkage region of fibroblast heparan sulphate

Lyon, Malcolm; Steward, William P.; Hampson, Ian N.; Gallagher, John T.

doi:10.1042/bj2420493

Cited by 42 publications

(26 citation statements)

References 23 publications

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“…5). An unmodified sequence of similar size was found in fibroblast HS (36), whereas heparin can be N-sulfated one to three disaccharides from the linkage sequence (34,37). Drosophila HS of lower molecular size than assumed here would display similar domain arrangement as in Fig.…”

Section: Discussionsupporting

confidence: 51%

Drosophila Heparan Sulfate, a Novel Design

Kusche-Gullberg

Nybakken

Perrimon

et al. 2012

Journal of Biological Chemistry

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Background: Heparan sulfate (HS) has important roles in cellular signaling that depend on the arrangement of sulfated domains. Results: Drosophila HS contains an essentially single sulfated domain. Conclusion:The domain organization of Drosophila HS is novel and differs from previously described HS structures. Significance: The findings may provide novel insight into structure-function relationships for HS.

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Section: Discussionsupporting

confidence: 51%

Drosophila Heparan Sulfate, a Novel Design

Kusche-Gullberg

Nybakken

Perrimon

et al. 2012

Journal of Biological Chemistry

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“…The fact that weak alkaline reduction, which failed to label proteoglycan side chains, provided reasonable labeling further indicates there to be little covalently attached oligopeptide. The linkage regions of cell and medium HSPGs were also within an N-sulfated domain, whereas in other cell types the HSPGs may have an extended N-acetylated sequence at this site (Lyon et al, 1987;Turnbull and Gallagher, 1991 ;Lindblom et al, 1991).…”

Section: Discussionmentioning

confidence: 99%

Differentially expressed patterns of glycosaminoglycan structure in heparan sulfate proteoglycans and free chains

Hovingh¹,

Piepkorn²,

Linker³

1993

European Journal of Biochemistry

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The metabolic relationships between heparan sulfate proteoglycans, free chains, and oligosaccharides in different cell locations were evaluated by comparing their glycosaminoglycan structure. Metabolically labeled heparan sulfate proteoglycans of BALBlc 3T3 cell layers and in conditioned medium were compared with the heparan sulfate free chains (modal mass = 10 kDa) and oligosaccharides (modal mass = 3 kDa) of the cells. Nonlytic, in situ digestion with heparitinase I indicated that 90% of proteoglycans, 70% of the free chains, and 20% of the oligosaccharides were enzyme accessible, but there was no evidence using competitive ligands for binding of the products to the cell surface via the glycosaminoglycan moieties. Structurally, the membrane proteoglycans were the most 0-lN-sulfated and yielded more tri-and tetra-sulfated di-and tetra-saccharides by nitrous acid degradation. In contrast, the side chains of medium proteoglycans were less sulfated and more polydisperse in mass, suggesting that most medium proteoglycans are not processed from membrane precursors. The heparan sulfate free chains were of lower mass, less sulfated, and more heterogeneous in distribution of the anionic groups than were proteoglycan side chains. Corroborating analytical heparitinase I digestion indicted that generation of di-and tetra-saccharides proportionately increased from membrane proteoglycan, to cell free chain, to medium proteoglycan categories. Because the structural patterns of the heparan sulfate free chains did not reveal a clear relationship with the side chains of the major proteoglycans, their origin was further probed by [3H]BH,-labeling of the reducing terminus under varying stringencies. The end-labeled residues obtained by nitrous or strong acid hydrolysis of the free chains showed insignificant amounts of galactose and xylose, but rather glucosamine N-sulfate and a residue likely generated from glucuronate. The effective labeling that was achieved with weak alkali indicated that covalent oligopeptide is not present. In summary, the heparan sulfate free chains, which in part are components of the cell surface, are of relatively low mass, are unassociated with covalent peptide, and most probably have a disaccharide motif of glucosamine N-sulfate and a uronate residue at the reducing end. Taken together, these observations suggest that the free chains originate by processing of precursor heparan sulfate proteoglycans on the cell surface via an endoglycosidase acting on an N-sulfated portion of the original polymer.

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“…These differences may conceivably be ascribed to effects of HS proteoglycan turnover, mediated by the endoglucuronidase, heparanase. The low molecular weight fractions III may, largely or in part, consist of chain fragments released from the peripheral portions of proteoglycan-associated longer chains by heparanase, and thus lack the predominantly N-acetylated saccharide regions proximal to the core protein(s) (51).…”

Section: Hs Chain Length-related Substitutionmentioning

confidence: 99%

Heparan Sulfate Domain Organization and Sulfation Modulate FGF-induced Cell Signaling

Jastrebova

Vanwildemeersch

Lindahl

et al. 2010

Journal of Biological Chemistry

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Heparan sulfates (HSs) modulate various developmental and homeostatic processes by binding to protein ligands. We have evaluated the structural characteristics of porcine HS in cellular signaling induced by basic fibroblast growth factor (FGF2), using CHO745 cells devoid of endogenous glycosaminoglycans as target. Markedly enhanced stimulation of cell signaling, measured as phosphorylation of ERK1/2 and protein kinase B, was only observed with the shortest HS chains isolated from liver, whereas the longer chains from either liver or intestine essentially prolonged duration of signals induced by FGF2 in the absence of polysaccharide. Structural analysis showed that contiguous sulfated domains were most abundant in the shortest HS chains and were more heavily sulfated in HS from liver than in HS from intestine. Moreover, the shortest chains from either source entered into ternary complexes with FGF2 and FGF receptor-1c more efficiently than the corresponding longer chains. In addition to authentic HSs, decasaccharide libraries generated by chemo-enzymatic modification of heparin were probed for effect on FGF2 signaling. Only the most highly sulfated decamers, previously found most efficient in ternary complex formation (Jastrebova, N., Vanwildemeersch, M., Rapraeger, A. C., Giménez-Gallego, G., Lindahl, U., and Spillmann, D. (2006) J. Biol. Chem. 281, 26884 -26892), promoted FGF2 cellular signaling as efficiently as short HS chains from liver. Together these results suggest that the effects of HS on FGF2 signaling are determined by both the structure of the highly sulfated domains and by the organization/availability of such domains within the HS chain. These findings underpin the need for regulation of HS biosynthesis in relation to control of growth factor-induced signaling pathways.

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Identification of an extended N-acetylated sequence adjacent to the protein-linkage region of fibroblast heparan sulphate

Cited by 42 publications

References 23 publications

Drosophila Heparan Sulfate, a Novel Design

Drosophila Heparan Sulfate, a Novel Design

Differentially expressed patterns of glycosaminoglycan structure in heparan sulfate proteoglycans and free chains

Heparan Sulfate Domain Organization and Sulfation Modulate FGF-induced Cell Signaling

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