Open AccessIn vitro assay for the Bacillus subtilis signal peptidase SipS: Systems for efficient in vitro transcription‐translation and processing of precursors of secreted proteins
Author(s) -
Vehmaanperä Jari,
Görner Ariane,
Venema Gerard,
Bron Sierd,
Dijl Jan Maarten
Publication year - 1993
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1993.tb06575.x
Subject(s) - bacillus subtilis , in vitro , signal peptidase , biology , biochemistry , transcription (linguistics) , protein biosynthesis , translation (biology) , signal peptide , bacteria , messenger rna , recombinant dna , genetics , gene , linguistics , philosophy
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.