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Probing substrate recognition of bacterial lipoprotein signal peptidase using FRET reporters
Author(s) -
Kitamura Seiya,
Wolan Dennis W.
Publication year - 2018
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.13155
Subject(s) - escherichia coli , biochemistry , chemistry , substrate (aquarium) , lipoprotein(a) , protease , förster resonance energy transfer , lipoprotein , biology , enzyme , gene , cholesterol , ecology , physics , quantum mechanics , fluorescence
Lipoprotein signal peptidase (Lsp) is a transmembrane aspartic acid protease with a pivotal role in the bacterial lipoprotein maturation pathway. Despite the universal use of Lsp across the Bacterial Kingdom and its potential as an antibiotic target, the substrate recognition patterns of Lsp are poorly understood. Here, we investigated the substrate recognition and biochemical properties of Lsp from Gram − ( Escherichia coli ) and Gram + ( Streptococcus pyogenes ) bacteria, using synthetic peptide‐based FRET reporters. Didecanoyl glycerol was found to be an optimal lipid length, and Lsp demonstrated exclusive enantio‐selectivity for the ( R )‐form of the diacylglycerol. Our study will facilitate the iterative optimization of in vitro Lsp assays, as well as provide the first chemical interrogation into the substrate scope of Lsp.