Activation of two new α(1,3)fucosyltransferase activities in Chinese hamster ovary cells by 5-azacytidine

Barry Potvin, Pamela Stanley

Research output: Contribution to journalArticlepeer-review

33 Scopus citations

Abstract

Several mammalian α(1,3)fucosyltransferases (α[1,3]Fuc-T) that synthesize carbohydrates containing α(1,3)fucosylated lactosamine units have been identified. Although Chinese hamster ovary (CHO) cells do not express α(1,3)Fuc-T activity, the rare mutants LEC11 and LEC12, isolated after mutagenesis or DNA transfection, each express an α(1,3)Fuc-T that may be distinguished by several criteria. Two new CHO mutants possessing α(1,3)Fuc-T activity (LEC29 and LEC30) have now been isolated after treatment of a CHO cell population with 5-azacytidine (5-AzaC), ethylnitrosourea (ENU), or 5-AzaC followed by N-methyl-N′-nitro-N-nitrosoguanidine (MNNG). Like LEC12, both mutants possess an N-ethylmaleimide-resistant α(1,3)Fuc-T activity that can utilize a variety of acceptors and both express the Lewis X (Lex) determinant {Galβ[1,4](Fucα[1,3])GlcNAcβ1} but not the sialylα(2,3)Lex determinant on cell-surface carbohydrates. However, LEC29 and LEC30 may be distinguished from LEC11 and LEC12, as well as from each other, on the basis of their unique patterns of lectin resistance and their abilities to bind the VIM-2 monoclonal antibody that recognizes carbohydrates terminating in NeuNAcα(2,3)Galβ(1,4)GlcNAcβ(1,3)Galβ(1,4)(Fucα[1,3]) GlcNAcβ and also by the different in vitro substrate specificities and kinetic properties of their respective α(1,3)Fuc-T activities. The combined data provide good evidence that the LEC29 and LEC30 α(1,3)Fuc-Ts are novel transferases encoded by distinct gene products.

Original languageEnglish (US)
Pages (from-to)989-1000
Number of pages12
JournalMolecular biology of the cell
Volume2
Issue number12
DOIs
StatePublished - Dec 1991

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Fingerprint

Dive into the research topics of 'Activation of two new α(1,3)fucosyltransferase activities in Chinese hamster ovary cells by 5-azacytidine'. Together they form a unique fingerprint.

Cite this