Abstract
UDP-GlcNAc:GlcNAc β1-2Manα1-6R (GlcNAc to Man) β1,6-N-acetylglucosaminyltransferase V (GlcNAc-T V) adds a GlcNAcβ1-6 branch to bi- and triantennaryN-glycans. An increase in this activity has been associated with cellular transformation, metastasis and differentiation. We have used synthetic substrate analogues to study the substrate specificity and inhibition of the partially purified enzyme from hamster kidney and of extracts from hen oviduct membranes and acute myeloid leukaemia leukocytes. All compounds with the minimum structure GlcNAcβ1-2Manα1-6Glc/Manβ-R were good substrates for GlcNAc-T V. The presence of structural elements other than the minimum trisaccharide structure affected GlcNAc-T V activity without being an absolute requirement for activity. Substrates with a biantennary structure were preferred over linear fragments of biantennary structures. Kinetic analysis showed that the 3-hydroxyl of the Manα1-3 residue and the 4-hydroxyl of the Manβ- residue of the Manα1-6(Manα1-3)Manβ-RN-glycan core are not essential for catalysis but influence substrate binding. GlcNAcβ1-2(4,6-di-O-methyl-)Manα1-6Glcβ-pnp was found to be an inhibitor of GlcNAc-T V from hamster kidney, hen oviduct microsomes and acute and chronic myeloid leukaemia leukocytes.
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Abbreviations
- all:
-
allyl
- AML:
-
acute myeloid leukaemia
- BSA:
-
bovine serum albumin
- CML:
-
chronic myelogenous leukaemia
- Gal:
-
G,d-galactose
- Glc:
-
d-glucose
- GlcNAc:
-
Gn,N-acetyl-d-glucosamine
- HPLC:
-
high performance liquid chromatography
- Man:
-
M,d-mannose
- mco:
-
8-methoxycarbonyl-octyl, (CH2)8COOCH3
- Me:
-
methyl
- MES:
-
2-(N-morpholino)ethanesulfonate
- oct:
-
octyl
- pnp:
-
p-nitrophenyl
- T:
-
transferase
References
Varki A (1993)Glycobiology 3:97–30.
Varki A (1992)Curr Opin Cell Biol 4:257–66.
Feizi T (1993)Curr Opin Struct Biol 3:701–10.
Dennis JW (1992) InCell Surface Carbohydrates and Cell Development, (Fukuda M, ed.) pp 161–94, Boca Raton, Florida: CRC Press, Inc.
Dennis JW, Laferte S (1988) InAltered Glycosylation in Tumor Cells (Reading CL, Hakomori S-I, Marcus DM, eds) pp. 257–67, New York, N.Y.: Alan R. Liss Inc.
Smets LA, Van Beek WP (1984)Biochim Biophys Acta 738:237–49.
Santer UV, DeSantis R, Hard KJ, Van-Kuik JA, Vliegenthart JFG, Won B, Glick MC (1989)Eur J Biochem 181:249–60.
Dennis JW, Laferte S, Waghorne C, Breitman ML, Kerbel RS (1987)Science 236:582–85.
Pierce M, Arango J (1986)J Biol Chem 261:10772–77.
Arango J, Pierce M (1988)J Cell Biochem 37:225–31.
Yamashita K, Tachibana Y, Ohkura T, Kobata A (1985)J Biol Chem 260:3963–69.
Palcic MM, Ripka J, Kaur KJ, Shoreibah M, Hindsgaul O, Pierce M (1990)J Biol Chem 265:6759–69.
Yousefi S, Higgins E, Daoling Z, Pollex-Krüger A, Hindsgaul O, Dennis JW (1991)J Biol Chem 266:1772–82.
Easton EW, Bolscher JGM, Van den Eijnden DH (1991)J Biol Chem 266:21674–80.
Dennis JW, Laferte S (1989)Cancer Res 49:945–50.
Easton EW, Blokland I, Geldof AA, Rao BR, Van den Eijnden DH (1992)Febs Lett 308:46–49.
Dennis JW, Kosh K, Bryce D-M, Breitman ML (1989)Oncogene 4:853–60.
Cummings RD, Kornfeld S (1984)J Biol Chem 259:6253–60.
van den Eijnden DH, Koenderman AHL, Schiphorst WECM (1988)J Biol Chem 263:12461–71.
Brockhausen I, Romero PA, Herscovics A (1991)Cancer Res 51:3136–42.
Heffernan M, Lotan R, Amos B, Palcic M, Takano R, Dennis JW (1993)J Biol Chem 268:1242–51.
Nakao H, Nishikawa A, Karasuno T, Nishiura T, Iida M, Kanayama Y, Yonezawa T, Tarui S, Taniguchi N (1990)Biochem Biophys Res Commun 172:1260–66.
Cummings RD, Trowbridge IS, Kornfeld S (1982)J Biol Chem 257:13421–27.
Hindsgaul O, Kaur KJ, Srivastava G, Blaszczyk-Thurin M, Crawley SC, Heerze LD, Palcic MM (1991)J Biol Chem 266:17858–62.
Shoreibah MG, Hindsgaul O, Pierce M (1992)J Biol Chem 267:2920–27.
Gu J, Nishikawa A, Tsuruoka N, Ohno M, Yamaguchi N, Kanagawa K, Taniguchi N (1993)J Biochem 113:614–19.
Shoreibah M, Perng GS, Adler B, Weinstein J, Basu R, Cupples R, Wen D, Browne JK, Buckhaults P, Fregien N, Pierce M (1993)J Biol Chem 268:15381–85.
Saito H, Nishikawa A, Gu JG, Ihara Y, Soejima H, Wada Y, Sekiya C, Niikawa N, Taniguchi N (1994)Biochem Biophys Res Commun 198:318–27.
Saito H, Nishikawa A, Gu JG, Ihara Y, Soejima H, Wada Y, Sekiya C, Niikawa N, Taniguchi N (1994)Biochem Biophys Res Commun 200:668–69.
Hindsgaul O, Tahir SH, Srivastava OP, Pierce M (1988)Carbohydrate Res 173:263–72.
Srivastava OP, Hindsgaul O, Shoreibah M, Pierce M (1988)Carbohydrate Res 179:137–61.
Crawley SC, Hindsgaul O, Alton G, Pierce M, Palcic MM (1990)Anal Biochem 185:112–17.
Brockhausen I, Grey AA, Pang H, Schachter H, Carver JP (1988)Glycoconjugate J 5:419–48.
Brockhausen I, Carver J, Schachter H (1988)Biochem Cell Biol 66:1134–51.
Kanie O, Crawley SC, Palcic MM, Hindsgaul O (1993)Carbohydr Res 243:139–64.
Khan SH, Crawley SC, Kanie O, Hindsgaul O (1993)J Biol Chem 268:2468–73.
Brockhausen I, Matta KL, Orr J, Schachter H (1985)Biochemistry 24:1866–74.
Khan SH, Abbas SA, Matta KL (1989)Carbohydr Res 193:125–39.
Brockhausen I, Möller G, Yang JM, Khan SH, Matta KL, Paulsen H, Grey AA, Shah RN, Schachter H (1992)Carbohydr Res 236:281–99.
Khan SH, Matta KL (1993)J Carbohydr Chem 12:335–48.
Khan SH, Abbas SA, Matta KL (1990)Carbohydr Res 205:385–97.
Khan SH, Matta KL (1993)Carbohydr Res 243:29–42.
Paulsen H, Meinjohanns E, Reck F, Brockhausen I (1993)Liebigs Ann Chem 737–750.
Brockhausen I, Kuhns W, Schachter H, Matta KL, Sutherland DR, Baker MA (1991)Cancer Res 51:1257–63.
Toki D, Granovsky MA, Reck F, Kuhns W, Baker MA, Matta KL, Brockhausen I (1994)Biochem Biophys Res Commun 198:417–23.
Brockhausen I, Hull E, Hindsgaul O, Schachter H, Shah RN, Michnick SW, Carver JP (1989)J Biol Chem. 264:11211–21.
Kobata A (1988)Biochimie 70:1575–85.
Yamashita K, Hitoi A, Taniguchi N, Yokosawa N, Tsukada Y, Kobata A (1983)Cancer Res 43:5059–63.
Baker MA, Taub RN, Kanani A, Brockhausen I, Hindenburg A (1985)Blood 66:1068–71.
Baker MA, Kanani A, Brockhausen I, Schachter H, Hindenburg A, Taub RN (1987)Cancer Res 47:2763–66.
Kanani A, Sutherland DR, Fibach E, Matta KL, Hindenburg A, Brockhausen I, Kuhns W, Taub RN, Van den Eijnden DH, Baker MA (1990)Cancer Res 50:5003–7.
Linker T, Crawley SC, Hindsgaul O (1993)Carbohydr Res 245:323–31.
Vella GJ, Paulsen H, Schachter H (1984)Can J Biochem Cell Biol 62:409–17.
Möller G, Reck F, Paulsen H, Kaur KJ, Sarkar M, Schachter H, Brockhausen I (1992)Glycoconjugate J 9:180–90.
Bendiak B, Schachter H (1987)J Biol Chem 262:5775–83.
Bendiak B, Schachter H (1987)J Biol Chem 262:5784–90.
Reck F, Meinjohanns E, Springer M, Wilkens R, Van Dorst JALM, Paulsen H, Möller G, Brockhausen I, Schachter H (1994)Glycoconjugate J 11:210–16.
Nishikawa Y, Pegg W, Paulsen H, Schachter H (1988)J Biol Chem 263:8270–81.
Kanie O, Crawley SC, Palcic M, Hindsgaul O (1994)Bioorg. Medicinal Chem 2:1231–41.
Gleeson PA, Schachter H (1983)J Biol Chem 258:6162–73.
Narasimhan S (1982)J Biol Chem 257:10235–42.
Khan SH, Duus JØ, Crawley SC, Palcic MM, Hindsgaul O (1994)Tetrahedron Asymmetry 5:2415–35.
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Brockhausen, I., Reck, F., Kuhns, W. et al. Substrate specificity and inhibition of UDP-GlcNAc:GlcNAcβ1-2Manα1-6R β1,6-N-acetylglucosaminyltransferase V using synthetic substrate analogues. Glycoconjugate J 12, 371–379 (1995). https://doi.org/10.1007/BF00731340
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DOI: https://doi.org/10.1007/BF00731340