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Title: An easy and fast method for the evaluation of Staphylococcus epidermidis type I signal peptidase inhibitors
Authors: Bockstael, Katrijn
Geukens, Nick
Rao, Smitha C
Herdewijn, Piet
Anné, Jozef
Van Aerschot, Arthur # ×
Issue Date: Aug-2009
Publisher: Elsevier science bv
Series Title: Journal of Microbiological Methods vol:78 issue:2 pages:231-237
Abstract: In the framework of the search for new antimicrobial therapies to combat resistant bacteria, the type I signal peptidase (SPase I) serves as a potentially interesting target for the development of antibacterials with a new mode of action. Bacterial SPases I play a key role in protein secretion as they are responsible for the cleavage of signal peptides from secreted proteins. For the Gram-positive Staphylococcus epidermidis, an important source of biofilm-associated infections, three putative SPases I (denoted Sip1, Sip2, Sip3) have been described, of which Sip1 lacks the catalytic lysine. Here, we report the in vitro activity of purified Sip2 and Sip3 using pre-SceD as a native preprotein substrate of S. epidermidis and in a FRET-based assay. For the latter, a novel internally quenched fluorescent peptide substrate based on the signal peptide sequence of this native preprotein was developed and specific cleavage of this synthetic fluorogenic peptide substrate was demonstrated. The latter in vitro assay represents a rapid and reliable tool in future research for the identification and validation of potential SPase I inhibitors.
URI: 
ISSN: 0167-7012
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Medicinal Chemistry (Rega Institute)
Laboratory of Molecular Bacteriology (Rega Institute)
Faculty of Science, Campus Kulak Kortrijk
× corresponding author
# (joint) last author

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