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
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.