title: In vitro assay for the Bacillus subtilis signal peptidase SipS: systems for efficient in vitro transcription-translation and processing of precursors of secreted proteins.
abstract: The signal peptidase (SPase) SipS of Bacillus subtilis is responsible for the processing of precursors of secreted proteins. It differs from the SPases of Gram-negative bacteria in structure and specificity. To assay the activity of SipS in vitro, two efficient transcription-translation systems for the synthesis of radio-labelled precursors were developed. The systems were completely derived from B. subtilis. Post-translational in vitro processing of pre-staphylokinase by SipS was demonstrated. SipS activity was stimulated in vitro by several non-ionic detergents, whereas it was not affected by a large variety of proteinase inhibitors. SipS shares the latter property with other SPases.
authors.affiliation: Department of Genetics, Center of Biological Sciences, Haren, The Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Vehmaanperä
authors.forename: A
authors.initials: A
authors.lastname: Görner
authors.forename: G
authors.initials: G
authors.lastname: Venema
authors.forename: S
authors.initials: S
authors.lastname: Bron
authors.forename: J M
authors.initials: JM
authors.lastname: van Dijl
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology*; D003902:Detergents / Q000494:pharmacology*; D010450:Endopeptidases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology*; D014176:Protein Biosynthesis; D012697:Serine Endopeptidases
chemical_list: D003902:Detergents; D008565:Membrane Proteins; D011480:Protease Inhibitors; D010450:Endopeptidases; D012697:Serine Endopeptidases; C026515:type I signal peptidase; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus