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Volume 150, Issue 5

Molecular and functional characterization of type I signal peptidase from Free

Abstract

is a facultative intracellular Gram-negative rod-shaped bacterium that has become an important cause of both community-acquired and nosocomial pneumonia. Numerous studies concerning the unravelling of the virulence mechanism of this important pathogen have been initiated. As evidence is now accumulating for the involvement of protein secretion systems in bacterial virulence in general, the type I signal peptidase (LepB) of was of particular interest. This endopeptidase plays an essential role in the processing of preproteins carrying a typical amino-terminal signal peptide, upon translocation across the cytoplasmic membrane. This paper reports the cloning and the transcriptional analysis of the gene encoding the type I signal peptidase (SPase). Reverse transcription PCR experiments showed clear expression when was grown both in culture medium, and also intracellularly in , a natural eukaryotic host of . In addition, LepB was shown to be encoded by a polycistronic mRNA transcript together with two other proteins, i.e. a LepA homologue and a ribonuclease III homologue. SPase activity of the LepB protein was demonstrated by complementation analysis in a temperature-sensitive mutant. Protein sequence and predicted membrane topology were compared to those of leader peptidases of other Gram-negative human pathogens. Most strikingly, a strictly conserved methionine residue in the substrate binding pocket was replaced by a leucine residue, which might influence substrate recognition. Finally it was shown by experiments that LepB is a target for (5,6)-6-[()-acetoxyethyl]-penem-3-carboxylate, a specific inhibitor of type I SPases.

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Keyword(s): SPase, signal peptidase
SGM
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2004-05-01
2024-04-19
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Molecular and functional characterization of type I signal peptidase from Legionella pneumophila
Microbiology 150, 1475 (2004); https://doi.org/10.1099/mic.0.26973-0
/content/journal/micro/10.1099/mic.0.26973-0
/content/journal/micro/10.1099/mic.0.26973-0
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