Soluble rat brain sialidase does not influence intracellular glycosylation of Golgi sialyltransferase or its constitutive glycoproteins.

Cytosol- and Golgi-enriched fractions were obtained from whole rat brain homogenates by density gradient centrifugation. Using a 4-methylumbelliferyl neuraminic acid substrate a soluble neural sialidase has been identified and characterised. The enzyme had optimal activity at pH 6.0 and a Km of 0.44 +/- 0.18 mM. The specific activity increased during postnatal development and this was in parallel with the described temporal changes in total brain neuraminic acid turnover. The potential of this enzyme to influence the intracellular processing of sialoglycoconjugates was also investigated. Cytosol fractions were incapable of releasing [14C]NeuNAC [( 14C]N-acetylneuramic acid) transferred to the glycoproteins of isolated Golgi membranes by their associated sialyltransferase. Further preincubation of Golgi membranes with soluble sialidase had no effect on their intrinsic sialyltransferase activity. These results demonstrate that no epigenetic regulation of processed sialoglycoconjugates occurs intracellularly and these finding are related to post-translational control of neural cell adhesion molecule (N-CAM) sialylation state.