A split ubiquitin system to reveal topology and released peptides of membrane proteins.

ABSTRACT:

Background

Membrane proteins define biological functions of membranes in cells. Extracellular peptides of transmembrane proteins receive signals from pathogens or environments, and are the major targets of drug developments. Despite of their essential roles, membrane proteins remain elusive in topological studies due to technique difficulties in their expressions and purifications.

Methods

First, the target gene is cloned into a destination vector to fuse with C terminal ubiquitin at the N or C terminus. Then, Cub vector with target gene and Nub^WT or Nub^G vectors are transformed into AP4 or AP5 yeast cells, respectively. After mating, the diploid cells are dipped onto selection medium to check the growth. Topology of the target protein is determined acc

SUBMITTER: Li QP 

PROVIDER: S-EPMC5581432 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature