title: Inhibition and resumption of processing of the staphylokinase in some Escherichia coli prlA suppressor mutants.
abstract: Escherichia coli strains carrying certain prlA mutations (prlA4 and prlA401) could not support the processing and export of staphylokinase, resulting in the accumulation of the precursor form under high-level synthesis conditions. In order to clarify the cause of the defect in the structure of staphylokinase, we constructed signal peptide mutations of sak which suppressed the processing defect in the prlA4 cells by site-directed mutagenesis. The processing defect was suppressed when glycine or asparagine was introduced in place of the serine residue at position 17 from the amino terminus of the signal peptide. Substitutions of glycine for the leucine residue at position 15 and for the serine residue at position 19 were also effective. Other mutations we constructed had no suppression activity. Taking account of the correlation between the suppression activity and the parameter value of each substituted amino acid for the beta-turn probability, we predict that the staphylokinase signal peptide requires a more bending structure at the end of the hydrophobic core to act efficiently in the prlA4 cells than in the prl+ cells and that a function of the PrlA protein necessary to recognize the staphylokinase signal peptide has become deficient through the prlA4 mutation.
authors.affiliation: Yakult Central Institute for Microbiological Research, Tokyo, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Iino
authors.forename: T
authors.initials: T
authors.lastname: Sako
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005998:Glycine; D007930:Leucine; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D021382:Protein Sorting Signals / Q000032:analysis; D013329:Structure-Activity Relationship
chemical_list: D021382:Protein Sorting Signals; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus; D007930:Leucine; D005998:Glycine