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J. Biol. Chem., Vol. 277, Issue 31, 28200-28211, August 2, 2002

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Dynamic Change of Neural Cell Adhesion Molecule Polysialylation on Human Neuroblastoma (IMR-32) and Rat Pheochromocytoma (PC-12) Cells during Growth and Differentiation^*

Geetha L. Poongodi, Nimmagadda Suresh, Subash C. B. Gopinath, Tschining Chang, Sadako Inoue, and Yasuo Inoue

From the Institute of Biological Chemistry, Academia Sinica, Taipei 115-29, Taiwan

Polysialic acid (PSA) is a regulatory epitope of neural cell adhesion molecule (NCAM) in homophilic adhesion of neural cells mediated by NCAM, is also known to be re-expressed in several human tumors, thus serves as an oncodevelopmental antigen. In this study, using a recently developed ultrasensitive chemical method in addition to immunochemical methods, growth stage-dependent and retinoic acid (RA)-induced differentiation-dependent changes of PSA expression in human neuroblastoma (IMR-32) and rat pheochromocytoma (PC-12) cells were analyzed both qualitatively and quantitatively. Both IMR-32 and PC-12 cells expressed PSA on NCAM, and the level of PSA expressed per unit weight of cells increased with post-inoculation incubation time. The most prominent feature was seen at the full confluence stage. RA induced neuronal differentiation in both IMR-32 and CP-12 cells that paralleled the change in the PSA level. Chemical analysis revealed the presence of NCAM glycoforms differing in the degree of polymerization (DP) of oligo/polysialyl chains, whose DP was smaller than 40. DP distribution of PSA was different between the cell lines and was changed by the growth stage and the RA treatment. Thus DP analysis of PSA is important in understanding both mechanism and biological significance of its regulated expression.

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