title: Effects of signal peptide mutations on processing of Bacillus stearothermophilus alpha-amylase in Escherichia coli.
abstract: Bacillus stearothermophilus alpha-amylase has a signal peptide typical for proteins exported by Gram-positive bacteria. There is only one signal peptidase processing site when the protein is exported from the original host, but when it is exported by Escherichia coli, two alternative sites are utilized. Site-directed mutagenesis was used to study the processing in E. coli. Processing sites for 13 B. stearothermophilus alpha-amylases carrying mutations in their signal peptide were determined. Processing of the signal peptide was remarkably tolerant to mutations, because switching between the alternative sites was possible. The length and the sequence of the region between the hydrophobic core and the cleavage site was crucial for determining the choice of the processing site. Some mutations more distal to the cleavage site also affected the site preference.
authors.affiliation: Department of Biochemistry, University of Turku, Finland.
authors.forename: I
authors.initials: I
authors.lastname: Suominen
authors.forename: P
authors.initials: P
authors.lastname: Meyer
authors.forename: C
authors.initials: C
authors.lastname: Tilgmann
authors.forename: T
authors.initials: T
authors.lastname: Glumoff
authors.forename: V
authors.initials: V
authors.lastname: Glumoff
authors.forename: J
authors.initials: J
authors.lastname: Käpylä
authors.forename: P
authors.initials: P
authors.lastname: Mäntsälä
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D002874:Chromosome Mapping; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D005798:Genes, Bacterial; D001411:Geobacillus stearothermophilus / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000235:genetics* / Q000378:metabolism*; D000516:alpha-Amylases / Q000235:genetics* / Q000378:metabolism*
chemical_list: D021382:Protein Sorting Signals; D000516:alpha-Amylases