ALPHA 1,6-LINKED FUCOSE AFFECTS THE EXPRESSION AND STABILITY OF POLYSIALIC ACID-CARRYING GLYCOPROTEINS IN CHINESE-HAMSTER OVARY CELLS

Citation
N. Kojima et al., ALPHA 1,6-LINKED FUCOSE AFFECTS THE EXPRESSION AND STABILITY OF POLYSIALIC ACID-CARRYING GLYCOPROTEINS IN CHINESE-HAMSTER OVARY CELLS, Journal of Biochemistry (Tokyo), 124(4), 1998, pp. 726-737
Citations number
45
Categorie Soggetti
Biology
ISSN journal
0021924X
Volume
124
Issue
4
Year of publication
1998
Pages
726 - 737
Database
ISI
SICI code
0021-924X(1998)124:4<726:A1FATE>2.0.ZU;2-S
Abstract
To determine the effect of alpha 1,6-linked fucose modification of N-g lycans 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 wi th 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 [SH]glucosamine for 48 h followed by immunopre cipitation with anti-PSA. monoclonal antibodies revealed that the amou nt of labeled PSA-carrying glycoproteins obtained from Lec13 cells was 10-times less than that from K1 cells, although the incorporation of [H-3]glucosamine into total extracts and NCAM was almost the same. In contrast, when cells were pulse labeled with [S-35]methionine followed by a Ih chase, there was not such a great difference in PSA-carrying protein synthesis between K1 and Lec13 cells. However, during a prolon ged chase period, PSA-carrying proteins rapidly decreased in Lec13 cel ls, whereas these in K1 cells did not change. The degradation of PSA-c arrying glycoproteins in Lec13 cells was partly prevented when the cel ls were grown in fucose-containing medium. Therefore, fucose modificat ion of core N-glycans may affect the efficient expression of PSAs thro ugh the intracellular stability of PSA-carrying glycoproteins.