In vitro insertion of leader peptidase into Escherichia coli membrane vesicles.
AUTOR(ES)
Moore, K E
RESUMO
Leader peptidase is an integral protein of the Escherichia coli cytoplasmic membrane whose topology is known. We have taken advantage of this knowledge and available mutants of this enzyme to develop a genetic test for a cell-free protein translocation reaction. We report that leader peptidase inserted into inverted plasma membrane vesicles in its correct transmembrane orientation. We have examined the in vitro membrane assembly characteristics of a variety of leader peptidase mutants and found that domains required for insertion in vivo are also necessary for insertion in vitro. These data demonstrate the physiological validity of the in vitro insertion reaction and strengthen the use of this in vitro protein translocation reaction for the dissection of this complex sorting pathway.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=211459Documentos Relacionados
- ATP is essential for protein translocation into Escherichia coli membrane vesicles.
- Tobramycin uptake in Escherichia coli membrane vesicles.
- Effects of nucleotides on ATP-dependent protein translocation into Escherichia coli membrane vesicles.
- Insertion of leader peptidase into the thylakoid membrane during synthesis in a chloroplast translation system
- Effect of diethylpyrocarbonate on lactose/proton symport in Escherichia coli membrane vesicles.