title: Role of the carboxy-terminal phenylalanine in the biogenesis of outer membrane protein PhoE of Escherichia coli K-12.
abstract: Most bacterial outer membrane proteins contain a phenylalanine at their C terminus. It has been shown that this residue has an important role in the efficient and correct assembly of PhoE protein into the Escherichia coli outer membrane, since its substitution or deletion resulted in the accumulation of trypsin-sensitive monomers of this normally trimeric protein. Here, the role of the C-terminal Phe in the assembly of PhoE was studied in further detail. Immunocytochemical labelling on ultrathin cryosections revealed that a mutant PhoE protein that lacks the C-terminal Phe accumulates in the periplasm. However, when the expression levels of the altered species were reduced, the efficiency of outer membrane incorporation was increased and the lethal effects were alleviated. The role of the C-terminal Phe in protein folding, trimerization and outer membrane incorporation was further studied in vitro. Deletion of this residue interfered with the efficiency of the formation of an assembly-competent folded monomer, and the stability of this PhoE form was affected. The in vitro trimerization and insertion into outer membranes were not affected by the mutation.
authors.affiliation: Department of Molecular Cell Biology and Institute of Biomembranes, Utrecht University, The Netherlands.
authors.forename: H
authors.initials: H
authors.lastname: de Cock
authors.forename: M
authors.initials: M
authors.lastname: Struyvé
authors.forename: M
authors.initials: M
authors.lastname: Kleerebezem
authors.forename: T
authors.initials: T
authors.lastname: van der Krift
authors.forename: J
authors.initials: J
authors.lastname: Tommassen
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002264:Carboxylic Acids; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D016296:Mutagenesis; D010649:Phenylalanine / Q000502:physiology*; D018272:Porins / Q000096:biosynthesis*; D017510:Protein Folding
chemical_list: D001426:Bacterial Proteins; D002264:Carboxylic Acids; D029968:Escherichia coli Proteins; D018272:Porins; C061279:PhoR protein, Bacteria; D010649:Phenylalanine; C082922:PhoE protein, E coli