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Binding, internalization, and degradation of antiproliferative heparan sulfate by human embryonic lung fibroblasts

1997; Wiley; Volume: 64; Issue: 4 Linguagem: Inglês

10.1002/(sici)1097-4644(19970315)64

1097-4644

Yolanda Arroyo‐Yanguas, Fang Cheng, Anders Isaksson, Lars‐Âke Fransson, Anders Malmström, Gunilla Westergren‐Thorsson,

Fibroblast Growth Factor Research

Journal of Cellular BiochemistryVolume 64, Issue 4 p. 595-604 Article Binding, internalization, and degradation of antiproliferative heparan sulfate by human embryonic lung fibroblasts Yolanda Arroyo-Yanguas, Yolanda Arroyo-Yanguas Department of Cell and Molecular Biology, Faculty of Medicine, Lund University, Lund, SwedenSearch for more papers by this authorFang Cheng, Fang Cheng Department of Cell and Molecular Biology, Faculty of Medicine, Lund University, Lund, SwedenSearch for more papers by this authorAnders Isaksson, Anders Isaksson Department of Cell and Molecular Biology, Faculty of Medicine, Lund University, Lund, SwedenSearch for more papers by this authorLars-Åke Fransson, Lars-Åke Fransson Department of Cell and Molecular Biology, Faculty of Medicine, Lund University, Lund, SwedenSearch for more papers by this authorAnders Malmström, Anders Malmström Department of Cell and Molecular Biology, Faculty of Medicine, Lund University, Lund, SwedenSearch for more papers by this authorGunilla Westergren-Thorsson, Corresponding Author Gunilla Westergren-Thorsson Department of Cell and Molecular Biology, Faculty of Medicine, Lund University, Lund, SwedenDepartment of Cell and Molecular Biology, Lund University, P.O. Box 94, S-221 00 Lund, SwedenSearch for more papers by this author Yolanda Arroyo-Yanguas, Yolanda Arroyo-Yanguas Department of Cell and Molecular Biology, Faculty of Medicine, Lund University, Lund, SwedenSearch for more papers by this authorFang Cheng, Fang Cheng Department of Cell and Molecular Biology, Faculty of Medicine, Lund University, Lund, SwedenSearch for more papers by this authorAnders Isaksson, Anders Isaksson Department of Cell and Molecular Biology, Faculty of Medicine, Lund University, Lund, SwedenSearch for more papers by this authorLars-Åke Fransson, Lars-Åke Fransson Department of Cell and Molecular Biology, Faculty of Medicine, Lund University, Lund, SwedenSearch for more papers by this authorAnders Malmström, Anders Malmström Department of Cell and Molecular Biology, Faculty of Medicine, Lund University, Lund, SwedenSearch for more papers by this authorGunilla Westergren-Thorsson, Corresponding Author Gunilla Westergren-Thorsson Department of Cell and Molecular Biology, Faculty of Medicine, Lund University, Lund, SwedenDepartment of Cell and Molecular Biology, Lund University, P.O. Box 94, S-221 00 Lund, SwedenSearch for more papers by this author First published: 07 December 1998 https://doi.org/10.1002/(SICI)1097-4644(19970315)64:4 3.0.CO;2-MCitations: 10AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat Abstract Binding, internalization, and degradation of 125I-labeled, antiproliferative, or nonantiproliferative heparan sulfate by human embryonic lung fibroblasts was investigated. Both L-iduronate-rich, antiproliferative heparan sulfate species as well as L-iduronate-poor, inactive ones were bound to trypsin-releasable, cell-surface sites. Both heparan sulfate types were bound with approximately the same affinity to one high-affinity site (Kd approximately 10−8 M) and to one (Kd approximately 10−6 M), respectively. Results of Hill-plot analysis suggested that the two sites are independent. Competition experiments with unlabeled glycosaminoglycans indicated that the binding sites had a selective specificity for sulfated, L-iduronate-rich heparan sulfate. Dermatan sulfate, which is also antiproliferative, was weakly bound to the cells. The antiproliferative effects of heparan and dermatan sulfate appeared to be additive. Hence, the two glycosaminoglycans probably exert their effect through different mechanisms. At concentrations above 5 μg/ml (approximately 10−7 M), heparan sulfate was taken up by human embryonic lung fibroblasts, suggesting that the low-affinity site represents an endocytosis receptor. The antiproliferative effect of L-iduronate-rich heparan sulfate species was also exerted at the same concentrations. The antiproliferative species was taken up to a greater degree than the inactive one, suggesting a requirement for internalization. However, competition experiments with dextran sulfate suggested that both the high-affinity and the low-affinity sites are involved in mediating the antiproliferative effect. Structural analysis of the inactive and active heparan sulphate preparations indicated that although sulphated L-iduronate appears essential for antiproliferative activity, it is not absolutely required for binding to the cells. Degradation of internalized heparan sulfate was analyzed by polyacrylamide gel electrophoresis using a sensitive detection technique. The inactive species was partially degraded, whereas the antiproliferative one was only marginally affected. J. Cell. Biochem. 64:595–604. © 1997 Wiley-Liss, Inc. References Belting M, Fransson L-Å (1993): The growth promoter spermine interacts specifically with dermatan sulfate regions that are rich in L-iduronic acid and possess antiproliferative activity. Glycoconjugate J 10: 453–460. 10.1007/BF00737966 CASPubMedWeb of Science®Google Scholar Belting M, Havsmark B, Jönsson M, Persson S, Fransson L-Å (1996): Heparan sulphate/heparin glycosaminoglycans with strong affinity for the growth-promoter spermine have high antiproliferative activity. Glycobiology 6: 121–129. 10.1093/glycob/6.2.121 CASPubMedWeb of Science®Google Scholar Bernfield M, Kokenyesi R, Kato M, Hinkes MT, Spring J, Gallo RL, Lose EJ (1992): Biology of the syndecans: A family of transmembrane heparan sulfate proteoglycans. Annu Rev Cell Biol 8: 365–393. 10.1146/annurev.cb.08.110192.002053 CASPubMedWeb of Science®Google Scholar Bidanset DJ, LeBaron R, Rosenberg L, Murphy-Ullrich JE, Hóók M (1992): Regulation of cell substrate adhesion: Effects of small galactosaminoglycan-containing proteoglycans. J Cell Biol 118: 1523–1531. 10.1083/jcb.118.6.1523 CASPubMedWeb of Science®Google Scholar Bradbury MG, Parish CR (1989): Receptors on lymphocytes for endogenous splenic glycosaminoglycans. Immunology 66: 546–553. CASPubMedWeb of Science®Google Scholar Busch SJ, Martin GA, Barnhart RL, Mano M, Cardin AD, Jackson RL (1992): Trans- repressor activity of nuclear glycosaminoglycans on fos and jun/AP-1 oncoproteinmediated transcription. J Cell Biol 116: 31–42. 10.1083/jcb.116.1.31 CASPubMedWeb of Science®Google Scholar Castellot JJ, Wong K, Herman B, Hoover RL, Albertini DF, Wright TC, Caleb BJ, Morrisk JK (1985): Binding and internalization of heparin by vascular smooth muscle cells. J Cell Physiol 124: 13–20. 10.1002/jcp.1041240104 CASPubMedWeb of Science®Google Scholar Cheng F, Yoshida K, Heinegård D, Fransson L-Å (1992): A new method for sequence analysis of glycosaminoglycans from heavily substituted proteoglycans reveals nonrandom positioning of 4-and 6–0-sulphated N-acetylgalactosamine in aggrecan-derived chondroitin sulphate. Glycobiology 2: 553–561. 10.1093/glycob/2.6.553 CASPubMedWeb of Science®Google Scholar Cheng F, Heinegård D, Malmström A, Schmidtchen A, Yoshida K, Fransson L-Å (1994): Patterns of uronosyl epimerization and 4-/6–0 sulphation in chondroitin/ dermatan sulphate from decorin and biglycan of various bovine tissue. Glycobiology 4: 685–696. 10.1093/glycob/4.5.685 CASPubMedWeb of Science®Google Scholar Fedarko NS, Conrad HE (1986): A unique heparan sulfate in the nuclei of hepatocytes: Structural changes with the growth state of the cells. J Cell Biol 102: 587–599. 10.1083/jcb.102.2.587 CASPubMedWeb of Science®Google Scholar Fransson L-Å (1987): Structure and function of cellassociated proteoglycans. Trends Biochem Sci 12: 406–411. 10.1016/0968-0004(87)90197-6 CASWeb of Science®Google Scholar Fransson L-Å Malmström A (1971): Structure of pig skin dermatan sulphate. 1. Distribution of D-glucuronic acid residues. Eur J Biochem 18: 422–430. 10.1111/j.1432-1033.1971.tb01259.x CASPubMedWeb of Science®Google Scholar Fransson L-Å Nieduszynski IA, Phelps CF, Sheehan JK (1979): Interactions between dermatan sulphate chains. III. Light-scattering and viscometry studies of selfassociation. Biochim Biophys Acta 586: 179–188. 10.1016/0304-4165(79)90416-1 CASWeb of Science®Google Scholar Fransson L-Å Sjöberg I, Havsmark B (1980a): Structural studies on heparan sulphates. Characterization of oligosaccharides; obtained by periodate oxidation and alkaline elimination. Eur J Biochem 106: 59–69. 10.1111/j.1432-1033.1980.tb05997.x CASPubMedWeb of Science®Google Scholar Fransson L-Å Nieduszynski IA, Sheehan JK (1980b): Interaction between heparan sulphate chains. I. A gel chromatographic, light-scattering and structural study of aggregating and non-aggregating chains. Biochim Biophys Acta 630: 287–300. 10.1016/0304-4165(80)90433-X CASPubMedWeb of Science®Google Scholar Fransson, L-Å Havsmark B, Silverberg B (1990): Amethod for the sequence analysis of dermatan sulphate. Biochem J 269: 381–388. 10.1042/bj2690381 CASPubMedWeb of Science®Google Scholar Gilles RJ, Didier N, Denton M (1986): Determination of cell number in monolayer cultures. Anal Biochem 159: 109–113. 10.1016/0003-2697(86)90314-3 CASPubMedWeb of Science®Google Scholar Glössl J, Schubert-Prinz R, Gregory JD, Damle SP, von Figura K, Kresse H (1983): Receptor-mediated endocytosis of proteoglycans by human fibroblasts involves recognition of the protein core. Biochem J 215: 295–301. 10.1042/bj2150295 CASPubMedWeb of Science®Google Scholar Hausser H, Ober B, Quentin-Hoffmann E, Schmidt B, Kresse H (1992): Endocytosis of different members of the small chondroitin/dermatan sulfate proteoglycan family. J Biol Chem 267: 11559–11564. CASPubMedWeb of Science®Google Scholar Herbert J-M, Maffrand J-P (1991): Effect of pentosan polysulphate, standard heparin and related compounds on protein kinase C activity. Biochim Biophys Acta 1091: 432–441. 10.1016/0167-4889(91)90211-F CASPubMedWeb of Science®Google Scholar Hiscock DRR, Yanagishita M, Hascall VC (1994): Nuclear localization of glycosaminoglycans in rat ovarian granulosa cells. J Biol Chem 269: 4539–4546. 10.1016/S0021-9258(17)41811-4 CASPubMedWeb of Science®Google Scholar Kjellén L, Lindahl U (1991): Proteoglycans: Structures and interactions. Annu Rev Biochem 60: 443–475. 10.1146/annurev.bi.60.070191.002303 CASPubMedWeb of Science®Google Scholar Kyhse-Andersen J (1984): Electroblotting of multiple gels: Asimple apparatus without buffer tank for rapidtransfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods 10: 203–209. 10.1016/0165-022X(84)90040-X CASPubMedWeb of Science®Google Scholar Lindblom A, Bengtsson-Olivecrona G, Fransson L-Å (1991): Domain structure of endothelial heparan sulphate. Biochem J 279: 821–829. 10.1042/bj2790821 CASPubMedWeb of Science®Google Scholar Malmström A, Carlstedt I, Åberg L, Fransson L-Å (1975): The copolymeric structure of dermatan sulphate produced by cultured human fibroblasts. Biochem J 151: 477–489. 10.1042/bj1510477 CASPubMedWeb of Science®Google Scholar Ottlinger ME, Pukac LA, Karnovsky MJ (1993): Heparin inhibits mitogen-activated protein kinase activation in intact rat vascular smooth muscle cells. J Biol Chem 268: 19173–19176. CASPubMedWeb of Science®Google Scholar Rapraeger AC, Krufka A, Olwin BB (1991): Requirements of heparan sulfate for bFGF-mediated fibroblast growth and myoblast differentiation. Science 252: 1705–1708. 10.1126/science.1646484 CASPubMedWeb of Science®Google Scholar Redini F, Moczar E, Antoine E, Poupon MF (1989): Binding and internalization of exogenous glycosaminoglycans in weakly and highly metastatic rhabdomyosarcoma cells. Biochem Biophys Actga 991: 359–366. 10.1016/0304-4165(89)90129-3 CASPubMedWeb of Science®Google Scholar Rodén L, Baker JR, Cifonelli JA, Mathews MB (1972): Isolation and characterization of connective tissue polysaccharides. Methods Enzymol 28: 73–139. 10.1016/0076-6879(72)28009-0 Google Scholar San Antonio JD, Lander AD, Wright TC, Karnovsky MJ (1992): Heparin inibits the attachment and growth of Balb/c-3T3 fibroblasts on collagen substrata. J Cell Physiol 150: 8–16. 10.1002/jcp.1041500103 CASPubMedWeb of Science®Google Scholar Schmidtchen A, Fransson L-Å (1992): Analysis of glycosaminoglycan chains from different proteoglycan populations in human embryonic skin fibroblasts. Eur J Biochem 208: 537–546. 10.1111/j.1432-1033.1992.tb17218.x CASPubMedWeb of Science®Google Scholar Schmidtchen A, Carlstedt I, Malmström A, Fransson L-Å (1990): Inventory of human skin fibroblast proteoglycan. Identification of multiple heparan and chondroitin/ dermatan sulphate proteoglycans. Biochem J 265: 289–300. 10.1042/bj2650289 CASPubMedWeb of Science®Google Scholar Shively JE, Conrad HE (1976): Formation of anhydrosugars in the chemical depolymerization of heparin. Biochemistry 15: 3932–3942. 10.1021/bi00663a005 CASPubMedWeb of Science®Google Scholar Turnbull JE, Gallagher JT (1988): Oligosaccharide mapping of heparan sulphate by polyacrylamide-gradient-gel electrophoresis and electrotransfer to nylon membrane. Biochem J 251: 597–608. 10.1042/bj2510597 CASPubMedWeb of Science®Google Scholar Vannucchi S, Pasquali F, Porciatti F, Chiarugi V, Magnelli L, Bianchini P (1988): Binding, internalization and degradation of heparin and heparin fragments by cultured endothelial cells. Thromb Res 49: 373–383. 10.1016/0049-3848(88)90240-X CASPubMedWeb of Science®Google Scholar Westergren-Thorsson G, Önnervik P-O, Fransson L-Å, Malmström A (1991): Proliferation of cultured fibroblasts is inhibited by L-iduronate-containing glycosaminoglycans. J Cell Physiol 147: 523–530. 10.1002/jcp.1041470319 CASPubMedWeb of Science®Google Scholar Westergren-Thorsson G, Persson S, Isaksson A, Önnervik P-O, Malmström A, Fransson L-Å (1993): L-iduronaterich glycosaminoglycans inhibit growth of normal fibroblasts independently of serum or added growth factors. Exp Cell Res 206: 93–99. 10.1006/excr.1993.1124 CASPubMedWeb of Science®Google Scholar Wright TC Jr, Pukac LA, Castellot JJ Jr, Karnovsky MJ, Levine RA, Kim-ParkH-Y, Campisi J (1989): Heparin suppresses the induction of c-fos and c-myc mRNA in murine fibroblasts by selective inhibition of a protein kinase C-dependent pathway. Proc Natl Acad Sci USA 86: 3199–3203. 10.1073/pnas.86.9.3199 CASPubMedWeb of Science®Google Scholar Yayon A, Klagsbrun M, Esko JD, Leder P, Ornitz DM (1991): Cell surface, heparin-like molecules are required for binding of basic fibroblast growth factor to its high affinity receptor. Cell 64: 841–848. 10.1016/0092-8674(91)90512-W CASPubMedWeb of Science®Google Scholar Citing Literature Volume64, Issue415 March 1997Pages 595-604 ReferencesRelatedInformation