Fine-structural changes in rat liver microsomes treated with phospholipase C.

Abstract

Phospholipase C hydrolyses 50% of the phospholipids of rat liver microsomes. Our previous observations suggest that the hydrolysis is of the outer leaflet of the membrane bilayer. In the present investigation the products of hydrolysis and effect of phospholipase C treatment on the morphology of total, rough and smooth microsomes were investigated. The only product of phospholipase C treatment of microsomes was diglyceride. After hydrolysis of half the phospholipid, the microsomes retained their vesicular structure; however, the average diameter of total and smooth microsomes was reduced by approximately half, and that of rough microsomes by a third. Profiles were observed that suggested that small vesicles may form by pinching off from large vesicles. About 10-15% of the phospholipase-treated microsomes had associated with them an amorphous droplet yielding a 'signet-ring'-like structure. The rest of the vesicle membrane retained a bilayer structure. About 10-15% of the treated vesicles adhered in groups of two or three. In these groups the shared membrane was trilaminar. Freeze-fracture replicas of phospholipase C-treated microsomes exhibited a similar morphology to untreated microsomes having both concave and convex surfaces, the former exhibiting a greater density of intramembranous particles. These observations suggest that microsomal vesicles remain closed after treatment with phospholipase C and that the bilayer structure is retained. However, there are morphological changes that are possibly related to maintaining the stability of the membranes in which the outer leaflet consists essentially of diglyceride.