DIFFERENTIAL ACTION OF BACTERIAL PHOSPHATIDYLINOSITOL–SPECIFIC PHOSPHOLIPASES C ON BOVINE ERYTHROCYTE MEMBRANE GHOST

DEEPALI K. KOTHEKAR; DEBJANI DASGUPTA

Volume : V, Issue : VII, July - 2015

DIFFERENTIAL ACTION OF BACTERIAL PHOSPHATIDYLINOSITOL–SPECIFIC PHOSPHOLIPASES C ON BOVINE ERYTHROCYTE MEMBRANE GHOST

Deepali K. Kothekar, Debjani Dasgupta

Abstract :

In recent years, much attention has been focused on identifying the glycosylphosphatidylinositol (GPI) anchors of eukaryotic memane proteins and understanding their role in protein function. Commercial preparation of phosphatidylinositol–specific phospholipase C (PIPLC) from Bacillus cereusis routinely used as a tool for releasing and identifying GPI–anchored proteins (GPI–AP). Here, we compared the action of PIPLC from Bacilluscereus(Bc–PIPLC)and Staphylococcus aureus (Sa–PIPLC) on bovine erythrocyte memane ghost (BEMG). BEMG treated with Bc–PIPLC or Sa–PIPLC showed dissimilar electrophoretic pattern of released GPI–APs.The result indicates that the resistance and susceptibility of GPI–APs to PIPLC depends on the source of enzyme and this aspect warrants serious consideration while using PIPLC as a tool for identification of GPI–APs. Further, combination of PIPLCs from two sources showed a cumulative effect in releasing GPI–APs suggesting it to be more efficient in identification of GPI–APs than the conventional use of a single PIPLC preparation.