title: The synthesis of alpha-amylase by rough and in vitro reconstituted rough endoplasmic reticulum derived from rat parotid gland.
abstract: The isolation of rough and smooth endoplasmic reticulum from rat parotid salivary gland is described. The rough membrane was stripped of its bound ribosomes using the KCl-puromycin method. Rough endoplasmic reticulum was reconstituted from stripped-rough membrane and polyribosomes. The reconstituted rough membrane resembled the native rough membrane in the following aspects: RNA/protein ratio, buoyant density in a continuous sucrose gradient and amino acid incorporation capacity. The in vitro synthesis of alpha-amylase by both rough and in vitro reconstituted rough membrane was demonstrated using SDS polyacrylamide gel electrophoresis. The reconstituted rough membrane could be restripped by KCl-puromycin. The in vitro synthesized alpha-amylase remained associated with the rough or the in vitro reconstituted rough membrane, even after these membranes were stripped of their bound ribosomes.
mesh_terms: D000681:Amylases / Q000096:biosynthesis*; D000818:Animals; D002458:Cell Fractionation; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D007930:Leucine / Q000378:metabolism; D008274:Magnesium / Q000494:pharmacology; D010306:Parotid Gland / Q000201:enzymology*; D010649:Phenylalanine / Q000378:metabolism; D051381:Rats; D012270:Ribosomes / Q000201:enzymology; D013347:Subcellular Fractions / Q000201:enzymology
chemical_list: D010649:Phenylalanine; D000681:Amylases; D007930:Leucine; D008274:Magnesium