Biosynthesis of lipase in the scutellum of maize kernel.
In the scutellum of maize kernel after imbibition, lipase activity increased rapidly, concomitant with the decrease in storage triacylglycerols. The enzyme activity peaked at day 6, but remained at the same level from day 6-10 when most of the triacylglycerols had been depleted. By in vitro translation with extracted RNAs followed by immunoprecipitation, and by resolving the translation products by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, lipase was found to be de novo synthesized in postgermination. The enzyme was synthesized by RNAs extracted from free polyribosomes and not from bound polyribosomes. Both in vitro and in vivo synthesized lipase had the same Mr of 65,000 as resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, as had the purified authentic enzyme; thus there was no appreciable co- or post-translational processing of the enzyme. Lipase-specific mRNA was present only between day 2-6 after imbibition. At day 6 when lipolysis was most active, more than 60% of the lipase activity was recovered in the lipid body fraction and specifically associated with the organelle membrane. From day 6-10, the lipase activity gradually shifted from the lipid body fraction to other subcellular fractions, including the 10,000 X g pellet, the 120,000 X g pellet, and the 120,000 X g supernatant. Lipase in these subcellular fractions was attributed to represent the enzyme associated with membrane ghosts of the lipid bodies which were fusing with the fragile cell vacuoles; such fusions were observed in situ by electron microscopy.
D004533:Egtazic Acid / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D008049:Lipase / Q000096:biosynthesis* / Q000235:genetics; D008055:Lipids / Q000032:analysis; D008066:Lipolysis; D008274:Magnesium / Q000378:metabolism; D008854:Microscopy, Electron; D014176:Protein Biosynthesis; D012313:RNA / Q000032:analysis; D012639:Seeds / Q000201:enzymology; D003313:Zea mays / Q000201:enzymology*
D008055:Lipids; D004533:Egtazic Acid; D012313:RNA; D008049:Lipase; D008274:Magnesium