RRC ID 39279
Author Kojima N, Tachida Y, Tsuji S.
Title Alpha 1,6-linked fucose affects the expression and stability of polysialic acid-carrying glycoproteins in Chinese hamster ovary cells.
Journal J Biochem
Abstract To determine the effect of alpha1,6-linked fucose modification of N-glycans on the expression of polysialic acids (PSAs), the expression of PSAs in a fucose-lacking mutant of Chinese hamster ovary (CHO) cells, Lec13, was compared with that in CHO K1 cells. PSA synthase activity in these cells and the antennary structures of N-glycans associated with the neural adhesion molecule (NCAM), which is a major PSA-carrying glycoprotein, did not differ between the two types of cells. Metabolic labeling of cells with [3H]glucosamine for 48 h followed by immunoprecipitation with anti-PSA monoclonal antibodies revealed that the amount of labeled PSA-carrying glycoproteins obtained from Lec13 cells was 10-times less than that from K1 cells, although the incorporation of [3H]glucosamine into total extracts and NCAM was almost the same. In contrast, when cells were pulse labeled with [35S]methionine followed by a 1 h chase, there was not such a great difference in PSA-carrying protein synthesis between K1 and Lec13 cells. However, during a prolonged chase period, PSA-carrying proteins rapidly decreased in Lec13 cells, whereas those in K1 cells did not change. The degradation of PSA-carrying glycoproteins in Lec13 cells was partly prevented when the cells were grown in fucose-containing medium. Therefore, fucose modification of core N-glycans may affect the efficient expression of PSAs through the intracellular stability of PSA-carrying glycoproteins.
Volume 124(4)
Pages 726-37
Published 1998-10-1
DOI 10.1093/oxfordjournals.jbchem.a022173
PMID 9756617
MeSH Animals Antibodies, Monoclonal CHO Cells Clone Cells Cricetinae Fucose / deficiency Fucose / metabolism* Glucosamine / metabolism Glycoproteins / biosynthesis Glycoproteins / metabolism* Kinetics Neural Cell Adhesion Molecules / biosynthesis* Sialic Acids / biosynthesis Sialic Acids / metabolism* Sialyltransferases / metabolism* Tritium
IF 2.476
Times Cited 8
Human and Animal Cells