title: The pediocin AcH precursor is biologically active.
abstract: The properties of the pediocin AcH precursor, prepediocin AcH, have been studied to gain insight into how producer cells may protect themselves from the activity of intracellular prebacteriocins. The native 62-amino-acid precursor and the 44-amino-acid mature species were expressed in Escherichia coli host strains that lack the leader peptide processing enzyme, PapD. Both forms inhibited the growth of the test bacterium Listeria innocua Lin11, indicating that the native precursor is biologically active. The two species also were synthesized in the context of maltose-binding protein chimeric proteins to facilitate the measurement of their relative specific activities. The chimeric form of the precursor was approximately 80% as active as the chimeric mature species. Of relevance to cell protection and pediocin AcH production, it was determined that the precursor is strongly susceptible to inactivation by reducing agents and to degradation by chymotrypsin and endogenous E. coli proteases. Taken together, the results indicate that the activity of prepediocin AcH may have to be controlled prior to secretion to prevent toxicity to the host. Perhaps producer cells avoid membrane damage by maintaining the precursor in a reduced inactive state or by degrading molecules whose secretion is delayed.
authors.affiliation: Department of Animal Science, University of Wyoming, Laramie, Wyoming 82071, USA. labcin@uwyo.edu
authors.forename: B
authors.initials: B
authors.lastname: Ray
authors.forename: R
authors.initials: R
authors.lastname: Schamber
authors.forename: K W
authors.initials: KW
authors.lastname: Miller
mesh_terms: D018528:ATP-Binding Cassette Transporters; D000595:Amino Acid Sequence; D001430:Bacteriocins / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism; D002918:Chymotrypsin / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins; D006094:Gram-Positive Bacteria / Q000187:drug effects* / Q000254:growth & development; D058619:Maltose-Binding Proteins; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins; D000070017:Pediocins; D010957:Plasmids / Q000235:genetics; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D018528:ATP-Binding Cassette Transporters; D001430:Bacteriocins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D000070017:Pediocins; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C461599:maltose transport system, E coli; C074288:pediocin PA-1; D010450:Endopeptidases; D002918:Chymotrypsin