Repair of BHK cell surface ganglioside GM3 after its degradation by extracellular sialidase

Abstract

Treatment of BHK fibroblasts with V. cholerae sialidase for 20 min caused the breakdown of about 70% of total cellular ganglioside GM3 and the production of an approximately equivalent amount of lactosylceramide. On removal of the enzyme, a slow resynthesis of GM3 from lactosylceramide was observed, equivalent to about 5-6%/h of the degraded GM3. Resynthesis of degraded surface ganglioside has not previously been observed, but its magnitude is similar to previous measurements of the rate of protein resialylation after sialidase treatment. This suggests that resialylation of both lipid and protein is limited by vesicular transport of plasma membrane components through the trans-Golgi network [TGN] where sialyltransferase is thought to be localized. In contrast, resynthesis of sphingomyelin which has been degraded at the cell surface by exogenous sphingomyelinase is about five times faster than resynthesis of GM3 and may involve non-vesicular transport of ceramide.

Key Words:

• BHK
• ganglioside GM3
• sialidase
• sphingomyelinase
• frans-Golgi network