Abstract
Novel glycoproteins carrying sialyl-LeA (SLeA) antigens (SL-GP) were isolated from ascites fluid from a patient with colorectal cancer by immunoaffinity chromatography. Their characteristics, including binding capacity to E-selectin, were investigated. SL-GP showed a typical mucin type amino acid composition in which Ser, Thr and Pro together accounted for greater than 50% of the total amino acid residues. A large amount of carbohydrate (about 80%) was present in SL-GP. The number of O-glycans carrying SLeA antigens comprised about 9% of the total number of O-glycosidic chains. SL-GP could bind to IL-1β treated HUVEC, and the binding was inhibited by anti-E-selectin and anti-SLeA monoclonal antibodies. The binding of colorectal cancer cells, LS 180, to HUVEC was assayed in the presence of SL-GP, oligosaccharides prepared from SL-GP and human milk SLeA hexasaccharide. SL-GP inhibited the binding most effectively, whereas equivalent amounts of the SL-GP oligosaccharides and milk SLeA hexasaccharide inhibited it only slightly. These results constitute direct evidence that a unique arrangement of SLeA antigens on the polypeptide chain, probably a cluster, is essential for the binding to E-selectin.
Similar content being viewed by others
References
Bevilacqua MP, Stengelin S, Gimbrone MA, Seed B (1989) Science 243: 1160–4.
Lasky LA (1992) Science 258: 964–9.
Varki A (1994) Proc Natl Acad Sci USA 91: 7390–7.
Lee YC (1992) FASEB J 6: 3193–200.
Nakada H, Numata Y, Inoue M, Tanaka N, Kitagawa H, Funakoshi I, Fukui S, Yamashina I (1991) J Biol Chem 266: 12402–5.
Nakada H, Inoue M, Numata Y, Tanaka N, Funakoshi I, Fukui S, Mellors A, Yamashina I (1993) Proc Natl Acad Sci USA 90: 2495–9.
Polly MJ, Phillips ML, Wayner E, Nudelman E, Singhal AK, Hakomori S, Paulson JC (1991) Proc Natl Acad Sci USA 88: 6224–8.
Tyrrell D, James P, Rao N, Foxall C, Abbas S, Dasgupta F, Nashed M, Hasegawa A, Kiso M, Asa D, Kidd J, Brandley BK (1991) Proc Natl Acad Sci USA 88: 10372–6.
Bevilacqua MP, Nelson RM (1992) J Clin Invest 91: 927–38.
Lasky LA, Singer MS, Dowbenko D, Imai Y, Henzel WJ, Grimley C, Fennie C, Gillett N, Watson SR, Rosen SD (1992) Cell 69: 927–38.
Norgard KE, Moore KL, Diaz S, Stults NL, Ushiyama S, McEver RP, Cummings RD, Varki A (1993) J Biol Chem 268: 12764–74.
Kitagawa H, Nakada H, Numata Y, Kurosaka A, Fukui S, Funakoshi I, Kawasaki T, Shimada K, Inagaki F, Yamashina I (1988) J Biochem 104: 591–4.
Kurosaka A, Kitagawa H, Fukui S, Numata Y, Nakada H, Funakoshi I, Kawasaki T, Ogawa T, Iijima H, Yamashina I (1988) J Biol Chem 263: 8724–6.
Numata Y, Nakada H, Fukui S, Kitagawa H, Ozaki K, Inoue M, Kawasaki T, Funakoshi I, Yamashina I (1990) Biochem Biophys Res Commun 170: 981–5.
Bevilacqua MP, Pober JS, Mendrick DL, Coltran RS, Gimbrone Jr, MA(1987) Proc Natl Acad Sci USA 84: 9238–42.
Katayama M, Handa M, Ambo H, Araki Y, Hirai S, Kato I, Kawai Y, Watanabe K, Ikeda Y (1992) J Immunol Methods 153: 41–8.
Klug TL, LeDonne NC, Greber TF, Zurawski Jr, VR (1988) Cancer Res 48: 1505–11.
Suzuki J, Kondo A, Kato I, Hase S, Ikenaka T (1991) Agric Biol Chem 55: 283–4.
Hara S, Takemori Y, Yamaguchi M, Nakamura M, Ohkura Y (1987) Anal Biochem 164: 138–45.
Carlson DM (1968) J Biol Chem 243: 616–26.
Laemmli UK (1970) Nature 227: 680–5.
Hunter WH (1966) Atomic Energy Commission Science Radio-active Pharmaceuticals USA Symp Ser 6: 728 (GA Andrews, RM Kniseley, HN Wagner Sr, eds.)
Byrd JC, Nardelli J, Siddiqui B, Kim YS (1988) Cancer Res 48: 6678–85.
Nardelli J, Byrd JC, Ho J, Fearney FJ, Tasman-Jones C, Kim YS (1988) Pancreas 3: 631–41.
Gum JR, Byrd JC, Hicks JW, Toribara NW, Lamport DTA, Kim YS (1989) J Biol Chem 264: 6480–7.
Levinovitz A, Muhlhoff J, Isenmann S, Vestweber D (1993) J Cell Biol 121: 449–59.
Gendler SJ, Lancaster CA, Taylor-Papadimitriou J, Duhig T, Peat N, Burchell J, Pemberton I, Lalani EN, Wilson D (1990) J Biol Chem 265: 15286–93.
Kitagawa H, Nakada H, Fukui S, Kawasaki T, Yamashina I (1991) Biochem Biophys Res Commun 178: 1429–36.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Inoue, M., Nakada, H., Oka, Y. et al. Novel glycoproteins inhibiting the binding of colorectal cancer cells to E-selectin. Glycoconj J 14, 147–153 (1997). https://doi.org/10.1023/A:1018529419094
Issue Date:
DOI: https://doi.org/10.1023/A:1018529419094