Glycosphingolipid Research Using Endoglycoceramidase

Abstract

Endoglycoceramidase (EGCase) cleaves the linkage between oligosaccharides and ceramides of various glycosphingolipids, leaving simultaneously both intact ceramides and oligosaccharides. Taking advantage of the specificity of EGCase, we have found it very useful in the structural studies of glycosphingolipids.A detergent was required for EGCase to express full activity, possibly due to their hydrophobic nature, and therefore EGCase cannot be used for research on live cells. Recently, activator proteins regarding the stimulation of EGCase activity in the absence of detergents were discovered. When using activator proteins in place of Triton X-100 for stimulating EGCase activity, it was also noted to cause no damage to intact cells. It is thus possible by activator proteins to elucidate the biological functions of endogenous glycosphingolipids in situ by EGCase.