Trafficking and localization studies of recombinant alpha1, 3-fucosyltransferase VI stably expressed in CHO cells

Glycobiology
L BorsigE G Berger

Abstract

Peripheral alpha1,3-fucosylation of glycans occurs by the action of either one of five different alpha1,3-fucosyltransferases (Fuc-Ts) cloned to date. Fuc-TVI is one of the alpha1,3-fucosyltransferases which is capable to synthesize selectin ligands. The major alpha1, 3-fucosyltransferase activity in human plasma is encoded by the gene for fucosyltransferase VI, which presumably originates from liver cells. While the sequence, chromosomal localization, and kinetic properties of Fuc-TVI are known, immunocytochemical localization and trafficking studies have been impossible because of the lack of specific antibodies. Here we report on the development and characterization of a peptide-specific polyclonal antiserum monospecific to Fuc-TVI and an antiserum to purified soluble recombinant Fuc-TVI crossreactive with Fuc-TIII and Fuc-TV. Both antisera were applied for immunodetection in stably transfected CHO cells expressing the full-length form of this enzyme (CHO clone 61/11). Fuc-TVI was found to be a resident protein of the Golgi apparatus. In addition, more than 30% of cell-associated and released enzyme activity was found in the medium. Maturation and release of Fuc-TVI was analyzed in metabolically labeled CHO 61/11 cells follo...Continue Reading

Citations

Jun 24, 2004·The Journal of Membrane Biology·W W Young
Dec 2, 1999·Biochimica Et Biophysica Acta·E StaudacherL März
Mar 5, 1999·Biochimica Et Biophysica Acta·H J MaccioniJ A Martina
Mar 12, 2002·Glycobiology·Eric G Berger
Jul 19, 2000·Proceedings of the National Academy of Sciences of the United States of America·S Schnyder-CandrianE G Berger
Jun 3, 2005·Nature Reviews. Drug Discovery·Danielle H Dube, Carolyn R Bertozzi
Aug 10, 2012·Organic & Biomolecular Chemistry·M Carmen GalanBeat Ernst
Oct 18, 2015·Cellular and Molecular Life Sciences : CMLS·Sakari KellokumpuTuomo Glumoff
Oct 6, 2000·The Journal of Biological Chemistry·J LiW W Young

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