Identification and Characterization of a Periplasmic Aminoacyl-phosphatidylglycerol Hydrolase Responsible for Pseudomonas aeruginosa Lipid Homeostasis*

Arendt, Wiebke; Groenewold, Maike K.; Hebecker, Stefanie; Dickschat, Jeroen S.; Moser, Jürgen

doi:10.1074/jbc.m113.482935

Cited by 34 publications

(43 citation statements)

References 57 publications

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“…Both of the latter enzymes are aa-PG hydrolases encoded in operon with aaPGS. 35,56 These hydrolases are involved in controlling the levels of aminoacylated lipids in the membrane (Fig. 6B), and both proteins were found to increase bacterial resistance to antimicrobial compounds.…”

Section: Homeostasis and Utilization Of Aa-pg In The Membranementioning

confidence: 99%

“…6B), and both proteins were found to increase bacterial resistance to antimicrobial compounds. 35,56,57 While the cellular localization of AhyD remains undefined, VirJ is known to be anchored to the periplasmic surface in the cytoplasmic membrane of the gram-negative bacterium P. aeruginosa. VirJ exhibits broad substrate specificity and can hydrolyse the Ala-, Gly-, and Lys- moieties of artificial substrates.…”

Section: Homeostasis and Utilization Of Aa-pg In The Membranementioning

confidence: 99%

“…VirJ exhibits broad substrate specificity and can hydrolyse the Ala-, Gly-, and Lys- moieties of artificial substrates. 56 …”

Section: Homeostasis and Utilization Of Aa-pg In The Membranementioning

confidence: 99%

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Deciphering the tRNA-dependent lipid aminoacylation systems in bacteria: Novel components and structural advances

Fields

Roy

2017

RNA Biology

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ABSTRACTtRNA-dependent addition of amino acids to lipids on the outer surface of the bacterial membrane results in decreased effectiveness of antimicrobials such as cationic antimicrobial peptides (CAMPs) that target the membrane, and increased virulence of several pathogenic species. After a brief introduction to CAMPs and the various bacterial resistance mechanisms used to counteract these compounds, this review focuses on recent advances in tRNA-dependent pathways for lipid modification in bacteria. Phenotypes associated with amino acid lipid modifications and regulation of their expression will also be discussed.

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Section: Homeostasis and Utilization Of Aa-pg In The Membranementioning

confidence: 99%

Section: Homeostasis and Utilization Of Aa-pg In The Membranementioning

confidence: 99%

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Deciphering the tRNA-dependent lipid aminoacylation systems in bacteria: Novel components and structural advances

Fields

Roy

2017

RNA Biology

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“…Recent results demonstrate that precise tuning of cellular A-PG and/or L-PG concentrations is fundamental for bacterial resistance (2). Furthermore, regulatory circuits including specific aminoacyl-phosphatidylglycerol (aa-PG) hydrolases have been described for P. aeruginosa and E. faecium (13,14).…”

mentioning

confidence: 99%

Structures of two bacterial resistance factors mediating tRNA-dependent aminoacylation of phosphatidylglycerol with lysine or alanine

Hebecker

Krausze

Hasenkampf

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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The cytoplasmic membrane is probably the most important physical barrier between microbes and the surrounding habitat. Aminoacylation of the polar head group of the phospholipid phosphatidylglycerol (PG) catalyzed by Ala-tRNA Ala -dependent alanyl-phosphatidylglycerol synthase (A-PGS) or by Lys-tRNA Lys -dependent lysyl-phosphatidylglycerol synthase (L-PGS) enables bacteria to cope with cationic peptides that are harmful to the integrity of the cell membrane. Accordingly, these synthases also have been designated as multiple peptide resistance factors (MprF). They consist of a separable C-terminal catalytic domain and an N-terminal transmembrane flippase domain. Here we present the X-ray crystallographic structure of the catalytic domain of A-PGS from the opportunistic human pathogen Pseudomonas aeruginosa. In parallel, the structure of the related lysyl-phosphatidylglycerol-specific L-PGS domain from Bacillus licheniformis in complex with the substrate analog L-lysine amide is presented. Both proteins reveal a continuous tunnel that allows the hydrophobic lipid substrate PG and the polar aminoacyl-tRNA substrate to access the catalytic site from opposite directions. Substrate recognition of A-PGS versus L-PGS was investigated using misacylated tRNA variants. The structural work presented here in combination with biochemical experiments using artificial tRNA or artificial lipid substrates reveals the tRNA acceptor stem, the aminoacyl moiety, and the polar head group of PG as the main determinants for substrate recognition. A mutagenesis approach yielded the complementary amino acid determinants of tRNA interaction. These results have broad implications for the design of L-PGS and A-PGS inhibitors that could render microbial pathogens more susceptible to antimicrobial compounds.A-PGS | L-PGS | MprF | structure | tRNA

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“…This homolog corresponds to the biosynthesis of alanyl-PG in response to acidification [23]. In addition, P. aeruginosa also has a pH-dependent promoter-controlled gene which codes for alanyl-PG hydrolase [24]. This pair of enzymes in P. aerugi nosa forms a closed circuit of forming and breaking alanylated PG.…”

Section: Resultsmentioning

confidence: 99%

Characterization of N-Succinylation of L-Lysylphosphatidylglycerol in Bacillus subtilis Using Tandem Mass Spectrometry

Atila

Katselis

Chumala

et al. 2016

J. Am. Soc. Mass Spectrom.

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Abstract. Phospholipids generally dominate in bacterial lipids. The negatively charged nature of phospholipids renders bacteria susceptible to cationic antibiotic peptides. In comparison with Gram-negative bacteria, Gram-positive bacteria in general have much less zwitterionic phosphatidylethanolamine. However, they are known for producing aminoacylated phosphatidylglycerol (PG), especially positively charged L-lysyl-PG, which is catalyzed by lysyl-PG synthase MprF, which appears to have a broad range of specificity for L-aminoacyl transfer RNAs. In addition, many Gram-positive bacteria also have a dlt-gene-coded D-alanylation pathway for lipoteichoic acids and wall teichoic acids covalently attached to a glycolipid or peptidoglycan. D-Alanylation also masks the dominant negative charge of the phosphate-rich polymers of teichoic acids. Using mass spectrometry, we have recently observed that precursor scans in negative mode for deprotonated amino acid fragments were most sensitive for ester-linked amino acids. Such a scan for precursors generating an m/z 145 lysyl anion revealed lysyl-PG as well as an additional species 100 m/z units greater than lysyl-PG. This unexpected species corresponded precisely to the expected mass of Nsuccinylated lysyl-PG. Tandem mass spectrometry revealed a precise match to the fragmentation pattern of this putative new species. PG, lysyl-PG, and N-succinyl-lysyl-PG may form a complete loop of charge reversal from -1 to +1 and then back to -1. Analogous charge reversal by N-succinylation of lysine residues in the bacterial as well as eukaryotic proteomes has been recently discovered as a major posttranslational modification. Such modification in bacterial lipids is possibly catalyzed by an enzyme homologous to the enzymes that modify lysine residues in proteins.

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Identification and Characterization of a Periplasmic Aminoacyl-phosphatidylglycerol Hydrolase Responsible for Pseudomonas aeruginosa Lipid Homeostasis*

Cited by 34 publications

References 57 publications

Deciphering the tRNA-dependent lipid aminoacylation systems in bacteria: Novel components and structural advances

Deciphering the tRNA-dependent lipid aminoacylation systems in bacteria: Novel components and structural advances

Structures of two bacterial resistance factors mediating tRNA-dependent aminoacylation of phosphatidylglycerol with lysine or alanine

Characterization of N-Succinylation of L-Lysylphosphatidylglycerol in Bacillus subtilis Using Tandem Mass Spectrometry

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