The synthesis of α-amylase by rough andin vitroreconstituted rough endoplasmic reticulum derived from rat parotid gland
✍ Scribed by Bernard Cwikel; Rachel Avner; Henryk H. Czosnek; Abraham A. Hochberg; Nathan Groot
- Publisher
- Springer
- Year
- 1976
- Tongue
- English
- Weight
- 582 KB
- Volume
- 2
- Category
- Article
- ISSN
- 0301-4851
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✦ Synopsis
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 a-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 a-amylase remained associated with the rough or the in vitro reconstituted rough membrane, even after these membranes were stripped of their bound ribosomes.