Compartmental Utilization of Carboxyl-14C-Tripalmitin by Tissue Homogenate of Pine Seeds

Abstract

A tissue homogenate of megagemetophyte of germinating seeds of Jeffrey pine (Pinus Jefferii Grev. and Balf.) was incubated with sonication-dispersed and albumin-carried ¹⁴C-tripalmitin in order to elucidate the sequential and quantitative role of cellular organelles in utilizing lipid reserve in seeds. After 5 minutes at 30 C, 25% of the tracer was localized in the fat body fraction, 9% in the pellet containing mitochondria and glyoxysomes, 14% in the supernatant, and 2% was found as CO₂. Radioactivity increased with time of incubation in the latter three fractions indicating the forward direction of utilization. Fat bodies contained mainly lipases and hydrolyzed the tracer to palmitate with diglyceride and monoglyceride as intermediates. About two-thirds of the palmitate had left the fat bodies in 5 minutes and entered the pellet fraction within which the tracer was distributed 1:2 in mitochondria and glyoxysomes, respectively. Longer incubation reduced the ratio to 1:3 while both organelles acquired more radioactive intermediates. Labeled acetyl-CoA and intermediate of β-oxidation were found in both organelle-containing fractions. The supernatant fraction contained radioactive diglycerides, monoglycerides, palmitate, sterol esters, and phospholipids, indicating lipase activity and direct utilization of fatty acid for the synthesis of sterol esters and polar lipids.