Phospholipid composition, phospholipase A 2 and sphingomyelinase activities in rat liver nuclear membrane and matrix

Abstract

Little data has been published regarding the presence of lipolytic—neutral sphingomyelinase and phospholipase A 2—activities in the nuclear matrix and membranes of cells. Considering the influence of phospholipids and the above enzymes on transcription and replication, the phospholipid composition and lipolytic activities of the nuclear matrix and membrane was determined. Pure nuclei from normal rat liver cells were isolated after nuclease digestion and extraction of the nuclear material with low and high ionic strengths, in presence of reducing agents. Phosphatidylcholine was the main phospholipid in the nuclear membranes (44% from the total phospholipids) whereas the amount of phosphatidylserine was highest in the nuclear matrix (45%). The specific activity of phospholipase A 2 in nuclear membranes was similar to that from liver plasma membranes and in hen erythrocyte nuclear membranes, (2.3 ± 0.6 nmol/mg/min, SEM, n = 8) while in the nuclear matrix it was 20 times higher. It did not show specificity towards phosphatidylcholine and phosphatidylethanolamine as substrates. A high sphingomyelinase activity in the nuclear matrix of rat liver was found (12.7 ± 2.1 nmol/mg/min, SEM, n = 8) and this activity was affected by reducing agents (dithiotreitol). Our results showed that phospholipids phosphatidylcholine and phosphatidylserine were constituent part of the nuclear structures as were the enzymes phospholipase A 2 and sphingomyelinase. This is consist with a role for lipids and their breakdown products in metabolic events within the nucleus.