Protein translocation into Escherichia coli membrane vesicles is inhibited by functional synthetic signal peptides.
A synthetic peptide corresponding to the signal sequence of wild type Escherichia coli lambda-receptor protein (LamB) inhibits in vitro translocation of precursors of both alkaline phosphatase and outer membrane protein A into E. coli membrane vesicles (half-maximal inhibition at 1-2 microM). By contrast, the inhibitory effect was nearly absent in a synthetic peptide corresponding to the signal sequence from a mutant strain that harbors a deletion mutation in the LamB signal region and displays an export-defective phenotype for this protein in vivo. Two peptides derived from pseudorevertant strains that arose from the deletion mutant and exported LamB in vivo were found to inhibit in vitro translocation with effectiveness that correlated with their in vivo export ability. Controls indicated that these synthetic signal peptides did not disrupt the E. coli membrane vesicles. These results can be interpreted to indicate that the presequences of exported proteins interact specifically with a receptor either in the E. coli inner membrane or in the cytoplasmic fraction. However, biophysical data for the family of signal peptides studied here reveal that they will spontaneously insert into a lipid membrane at concentrations comparable to those that cause inhibition. Hence, an indirect effect mediated by the lipid bilayer of the membrane must be considered.
D000469:Alkaline Phosphatase / Q000378:metabolism; D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins / Q000378:metabolism*; D004792:Enzyme Precursors / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D008566:Membranes / Q000378:metabolism; D009943:Organophosphorus Compounds / Q000378:metabolism; D018272:Porins; D021382:Protein Sorting Signals / Q000494:pharmacology*; D011991:Receptors, Virus / Q000378:metabolism
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004792:Enzyme Precursors; D009943:Organophosphorus Compounds; D018272:Porins; D021382:Protein Sorting Signals; D011991:Receptors, Virus; C016532:maltoporins; C008705:octamethyl pyrophosphoramide; D000469:Alkaline Phosphatase