Preparation of chondroitin sulfate libraries containing disulfated disaccharide units and inhibition of thrombin by these chondroitin sulfates
Tóm tắt
Chondroitin sulfate (CS) containing GlcA-GalNAc(4,6-SO4) (E unit) and CS containing GlcA(2SO4)-GalNAc(6SO4) (D unit) have been implicated in various physiological functions. However, it has been poorly understood how the structure and contents of disulfated disaccharide units in CS contribute to these functions. We prepared CS libraries containing E unit or D unit in various proportions by in vitro enzymatic reactions using recombinant GalNAc 4-sulfate 6-O-sulfotransferase and uronosyl 2-O-sulfotransferase, and examined their inhibitory activity toward thrombin. The in vitro sulfated CSs containing disulfated disaccharide units showed concentration-dependent direct inhibition of thrombin when the proportion of E unit or D unit in the CSs was above 15–17%. The CSs containing both E unit and D unit exhibited higher inhibitory activity toward thrombin than the CSs containing either E unit or D unit alone, if the proportion of the total disulfated disaccharide units of these CSs was comparable. The thrombin-catalyzed degradation of fibrinogen, a physiological substrate for thrombin, was also inhibited by the CS containing both E unit and D unit. These observations indicate that the enzymatically prepared CS libraries containing various amounts of disulfated disaccharide units appear to be useful for elucidating the physiological function of disulfated disaccharide units in CS.
Tài liệu tham khảo
Kawai, Y., Seno, N., Anno, K.: Chondroitin polysulfate of squid cartilage. J. Biochem. 60, 317–321 (1966)
Suzuki, S., Saito, H., Yamagata, T., Anno, K., Seno, N., Kawai, Y., Furuhashi, T.: Formation of three types of disulfated disaccharides from chondroitin sulfates by chondroitinase digestion. J. Biol. Chem. 243, 1543–1550 (1968)
Stevens, R.L., Adachi, R.: Protease-proteoglycan complexes of mouse and human mast cells and importance of their beta-tryptase-heparin complexes in inflammation and innate immunity. Immunol. Rev. 217, 155–167 (2007)
Clement, A.M., Sugahara, K., Faissner, A.: Chondroitin sulfate E promotes neurite outgrowth of rat embryonic day 18 hippocampal neurons. Neurosci. Lett. 269, 125–128 (1999)
Tully, S.E., Mabon, R., Gama, C.I., Tsai, S.M., Liu, X., Hsieh-Wilson, L.C.: A chondroitin sulfate small molecule that stimulates neuronal growth. J. Am. Chem. Soc. 126, 7736–7737 (2004)
Kawashima, H., Hirose, M., Hirose, J., Nagakubo, D., Plaas, A.H.K., Miyasaka, M.: Binding of a Large Chondroitin Sulfate/Dermatan Sulfate Proteoglycan, Versican, to L-selectin, P-selectin, and CD44. J. Biol. Chem. 275, 35448–35456 (2000)
Hirose, J., Kawashima, H., Yoshie, O., Tashiro, K., Miyasaka, M.: Versican interacts with chemokines and modulates cellular responses. J. Biol. Chem. 276, 5228–5234 (2001)
Ueoka, C., Kaneda, N., Okazaki, I., Nadanaka, S., Muramatsu, T., Sugahara, K.: Neuronal cell adhesion, mediated by the heparin-binding neuroregulatory factor midkine, is specifically inhibited by chondroitin sulfate E. Structural ans functional implications of the over-sulfated chondroitin sulfate. J. Biol. Chem. 275, 37407–37413 (2000)
Deepa, S.S., Umehara, Y., Higashiyama, S., Itoh, N., Sugahara, K.: Specific molecular interactions of oversulfated chondroitin sulfate E with various heparin-binding growth factors. Implications as a physiological binding partner in the brain and other tissues. J. Biol. Chem. 277, 43707–43716 (2002)
Asada, M., Shinomiya, M., Suzuki, M., Honda, E., Sugimoto, R., Ikekita, M., Imamura, T.: Glycosaminoglycan affinity of the complete fibroblast growth factor family. Biochim. Biophys. Acta. 1790, 40–48 (2009)
Bergefall, K., Trybala, E., Johansson, M., Uyama, T., Naito, S., Yamada, S., Kitagawa, H., Sugahara, K., Bergström, T.: Chondroitin sulfate characterized by the E-disaccharide unit is a potent inhibitor of herpes simplex virus infectivity and provides the virus binding sites on gro2C cells. J. Biol. Chem. 280, 32193–32199 (2005)
Uyama, T., Ishida, M., Izumikawa, T., Trybala, E., Tufaro, F., Bergström, T., Sugahara, K., Kitagawa, H.: Chondroitin 4-O-sulfotransferase-1 regulates E disaccharide expression of chondroitin sulfate required for herpes simplex virus infectivity. J. Biol. Chem. 281, 38668–38674 (2006)
Li, F., Ten Dam, G.B., Murugan, S., Yamada, S., Hashiguchi, T., Mizumoto, S., Oguri, K., Okayama, M., van Kuppevelt, T.H., Sugahara, K.: Involvement of highly sulfated chondroitin sulfate in the metastasis of the Lewis lung carcinoma cells. J. Biol. Chem. 283, 34294–34304 (2008)
Basappa, Murugan, S., Sugahara, K.N., Lee, C.M., ten Dam, G.B., van Kuppevelt, T.H., Miyasaka, M., Yamada, S., Sugahara, K.: Involvement of chondroitin sulfate E in the liver tumor focal formation of murine osteosarcoma cells. Glycobiology 19, 735–742 (2009)
Ishii, M., Maeda, N.: Oversulfated chondroitin sulfate plays critical roles in the neuronal migration in the cerebral cortex. J. Biol. Chem. 283, 32610–32620 (2008)
Ra, H.J., Harju-Baker, S., Zhang, F., Linhardt, R.J., Wilson, C.L., Parks, W.C.: Control of promatrilysin (MMP7) activation and substrate-specific activity by sulfated glycosaminoglycans. J. Biol. Chem. 284, 27924–27932 (2009)
Clement, A.M., Nadanaka, S., Masayama, K., Mandl, C., Sugahara, K., Faissner, A.: The DSD-1 carbohydrate epitope depends on sulfation, correlates with chondroitin sulfate D Motifs, and is sufficient to promote neurite outgrowth. J. Biol. Chem. 273, 28444–18453 (1998)
Maeda, N., He, J., Yajima, Y., Mikami, T., Sugahara, K., Yabe, T.: Heterogeneity of the Chondroitin Sulfate Portion of Phosphacan/6B4 Proteoglycan Regulates Its Binding Affinity for Pleiotrophin/Heparin Binding Growth-associated Molecule. J. Biol. Chem. 278, 35805–25811 (2003)
Habuchi, O., Sugiura, K., Kawai, N.: Glucose branches in chondroitin sulfates from squid cartilage. J. Biol. Chem. 252, 4570–4576 (1977)
Kinoshita, A., Yamada, S., Haslam, S.M., Morris, H.R., Dell, A., Sugahara, K.: Novel tetrasaccharides isolated from squid cartilage chondroitin sulfate E contain unusual sulfated disaccharide units GlcA(3-O-sulfate)beta1-3GalNAc(6-O-sulfate) or GlcA(3-O-sulfate)beta1-3GalNAc. J. Biol. Chem. 272, 19656–19665 (1997)
Ito, Y., Habuchi, O.: Purification and characterization of N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase from the squid cartilage. J. Biol. Chem. 275, 34728–34736 (2000)
Ohtake, S., Ito, Y., Fukuta, M., Habuchi, O.: Human N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase cDNA is related to human B cell recombination activating gene-associated gene. J. Biol. Chem. 276, 43894–43900 (2001)
Yamaguchi, T., Ohtake, S., Kimata, K., Habuchi, O.: Molecular cloning of squid N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase and synthesis of a unique chondroitin sulfate containing E-D hybrid tetrasaccharide structure by the recombinant enzyme. Glycobiology 17, 1365–1376 (2007)
Habuchi, O., Moroi, R., Ohtake, S.: Emzymatic synthesis of chondroitin sulfate E by N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase purified from the squid cartilage. Anal. Biochem. 310, 129–136 (2002)
Kobayashi, M., Sugumaran, G., Liu, J., Shworak, N.W., Silbert, J.E., Rosenberg, R.D.: Molecular cloning and characterization of a human uronyl 2-sulfotransferase that sulfates iduronyl and glucuronyl residues in dermatan/chondroitin sulfate. J. Biol. Chem. 274, 10474–10480 (1999)
Ohtake, S., Kimata, K., Habuchi, O.: Recognition of sulfation pattern of chondroitin sulfate by uronosyl 2-O-sulfotransferase. J. Biol. Chem. 280, 39115–39123 (2005)
Delfert, D.M., Conrad, H.E.: Preparation and high-performance liquid chromatography of 3′-phosphoadenosine-5′-phospho[35 S]sulfate with a predetermined specific activity. Anal. Biochem. 148, 303–310 (1985)
Habuchi, O., Matsui, Y., Kotoya, Y., Aoyama, Y., Yasuda, Y., Noda, M.: Purification of chondroitin 6-sulfotransferase secreted from cultured chick embryo chondrocytes. J. Biol. Chem. 268, 21968–21974 (1993)
Toyoda, H., Kinoshita-Toyoda, A., Selleck, S.B.: Structural analysis of glycosaminoglycans in drosophila and caenorhabditis elegans and demonstration that tout-velu, a drosophila gene related to EXT tumor suppressors, affects heparan sulfate in vivo. J. Biol. Chem. 275, 2269–2275 (2000)
Henry, B.L., Monien, B.H., Bock, P.E., Desai, U.R.: A novel allosteric pathway of thrombin inhibition. EXOSITE II MEDIATED POTENT INHIBITION OF THROMBIN BY CHEMO-ENZYMATIC, SULFATED DEHYDROPOLYMERS OF 4-HYDROXYCINNAMIC ACIDS. J. Biol. Chem. 282, 31891–31899 (2007)
Vigdorovich, V., Miller, D., Strong, R.K.: Ability of hyaluronidase 2 to degrade extracellular hyaluronan is not required for its function as a receptor for jaagsiekte sheep retrovirus. J. Virol. 81, 3124–3129 (2007)
Esmon, C.T.: Cell mediated events that control blood coagulations and vascular injury. Annu. Rev. Cell Biol. 9, 1–26 (1993)
Conrad, H.E.: Heparin-biding proteins. Academic, London (1998)
Bourin, M.-C., Lindahl, U.: Functional role of the polysaccharide component of rabbit thrombomodulin proteoglycan Effects on inactivation of thrombin by antithrombin, cleavage of fibrinogen by thrombin and thrombin-catalysed activation of Factor V. Biochem. J. 270, 419–425 (1990)
Bourin, M.-C., Lundgren-Akerlund, E., Lindahl, U.: Isolation and characterization of the glycosaminoglycan component of rabbit thrombomodulin proteoglycan. J. Biol. Chem. 265, 15424–15431 (1990)
Pereira, M.S., Mulloy, B., Mourão, P.A.S.: Structure and anticoagulant activity of sulfated fucans: COMPARISON BETWEEN THE REGULAR, REPETITIVE AND LINEAR FUCANS FROM ECHINODERMS WITH THE MORE HETEROGENEOUS AND BRANCHED POLYMERS FROM BROWN ALGAE. J. Biol. Chem. 274, 7656–7667 (1999)