A cell-free translocation system using extracts of cultured insect cells to yield functional membrane proteins.

ABSTRACT: Cell-free protein synthesis is a powerful method to explore the structure and function of membrane proteins and to analyze the targeting and translocation of proteins across the ER membrane. Developing a cell-free system based on cultured cells for the synthesis of membrane proteins could provide a highly reproducible alternative to the use of tissues from living animals. We isolated Sf21 microsomes from cultured insect cells by a simplified isolation procedure and evaluated the performance of the translocation system in combination with a cell-free translation system originating from the same source. The isolated microsomes contained the basic translocation machinery for polytopic membrane proteins including SRP-dependent targeting components, translocation channel (translocon)-dependent

SUBMITTER: Ezure T 

PROVIDER: S-EPMC4259328 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature