Neisseria gonorrhoeae prepilin export studied in Escherichia coli

摘要: The pilE gene of Neisseria gonorrhoeae MS11 and a series pilE-phoA fusions were expressed in Escherichia coli. PhoA hybrid proteins shown to be located the membrane fraction cells, prepilin product was exclusively cytoplasmic membrane. Analysis prepilin-PhoA hybrids showed that first 20 residues can function as an efficient export (signal) sequence. This segment includes unbroken sequence 8 hydrophobic or neutral form N-terminal half 16-residue region prepilin. Neither nor processed by E. coli leader peptidase despite presence two consensus cleavage sites for this enzyme just after region. Comparisons specific molecular activities four analysis their susceptibility proteolysis trypsin proteinase K spheroplasts allow us propose models topology bulk evidence supports simplest models, which is anchored solely domain, with extreme N terminus facing cytoplasm longer C periplasm.