Detergent-accelerated hydrolysis of bacterial endotoxins and determination of the anomeric configuration of the glycosyl phosphate present in the “Isolated lipid A” fragment of the Bordetella pertussis endotoxin

Caroff, Martine; Tacken, Angelika; Szabó, Ladislas

doi:10.1016/0008-6215(88)84149-1

Cited by 221 publications

(213 citation statements)

References 27 publications

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“…Our assay is based on the observation that the Kdo-lipid A linkage (Fig. 2) is cleaved selectively by pH 4.5 hydrolysis at 100°C in the presence of SDS without measurable loss of ester-linked fatty acids or of the anomeric phosphate (34)(35)(36). The hydrolysis procedure can be applied directly to radioactive cells or to isolated membrane fractions from a sucrose gradient without first extracting LPS with phenol-containing solvents.…”

Section: Discussionmentioning

confidence: 99%

Function of Escherichia coli MsbA, an Essential ABC Family Transporter, in Lipid A and Phospholipid Biosynthesis

Zhou¹,

White²,

Polissi³

et al. 1998

Journal of Biological Chemistry

326

339

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Section: Discussionmentioning

confidence: 99%

Function of Escherichia coli MsbA, an Essential ABC Family Transporter, in Lipid A and Phospholipid Biosynthesis

Zhou¹,

White²,

Polissi³

et al. 1998

Journal of Biological Chemistry

326

339

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“…This LPS did not activate the NF-κB-dependent luciferase reported gene in HEK293 cells transiently transfected with human TLR2 (59). Lipid A was obtained by acid hydrolysis (60).…”

Section: Methodsmentioning

confidence: 99%

Deciphering tissue-induced Klebsiella pneumoniae lipid A structure

Llobet

Martínez-Moliner

Moranta

et al. 2015

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

134

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The outcome of an infection depends on host recognition of the pathogen, hence leading to the activation of signaling pathways controlling defense responses. A long-held belief is that the modification of the lipid A moiety of the lipopolysaccharide could help Gram-negative pathogens to evade innate immunity. However, direct evidence that this happens in vivo is lacking. Here we report the lipid A expressed in the tissues of infected mice by the human pathogen Klebsiella pneumoniae. Our findings demonstrate that Klebsiella remodels its lipid A in a tissue-dependent manner. Lipid A species found in the lungs are consistent with a 2-hydroxyacyl-modified lipid A dependent on the PhoPQ-regulated oxygenase LpxO. The in vivo lipid A pattern is lost in minimally passaged bacteria isolated from the tissues. LpxO-dependent modification reduces the activation of inflammatory responses and mediates resistance to antimicrobial peptides. An lpxO mutant is attenuated in vivo thereby highlighting the importance of this lipid A modification in Klebsiella infection biology. Colistin, one of the last options to treat multidrug-resistant Klebsiella infections, triggers the in vivo lipid A pattern. Moreover, colistin-resistant isolates already express the in vivo lipid A pattern. In these isolates, LpxO-dependent lipid A modification mediates resistance to colistin. Deciphering the lipid A expressed in vivo opens the possibility of designing novel therapeutics targeting the enzymes responsible for the in vivo lipid A pattern.

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“…Fn and Yp LPS were extracted using the hot phenol/water extraction method as previously described [26]. Lipid A was isolated after LPS was treated with RNase A, DNase I, and proteinase K by the method of Caroff et al [27].…”

Section: Methodsmentioning

confidence: 99%

Automated Lipid A Structure Assignment from Hierarchical Tandem Mass Spectrometry Data

Ting

Shaffer

Jones

et al. 2011

J. Am. Soc. Mass Spectrom.

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Infusion-based electrospray ionization (ESI) coupled to multiple-stage tandem mass spectrometry (MS n ) is a standard methodology for investigating lipid A structural diversity (Shaffer et al. J. Am. Soc. Mass. Spectrom. 18(6), [1080][1081][1082][1083][1084][1085][1086][1087][1088][1089][1090][1091][1092] 2007). Annotation of these MS n spectra, however, has remained a manual, expert-driven process. In order to keep up with the data acquisition rates of modern instruments, we devised a computational method to annotate lipid A MS n spectra rapidly and automatically, which we refer to as hierarchical tandem mass spectrometry (HiTMS) algorithm. As a first-pass tool, HiTMS aids expert interpretation of lipid A MS n data by providing the analyst with a set of candidate structures that may then be confirmed or rejected. HiTMS deciphers the signature ions (e.g., A-, Y-, and Z-type ions) and neutral losses of MS n spectra using a species-specific library based on general prior structural knowledge of the given lipid A species under investigation. Candidates are selected by calculating the correlation between theoretical and acquired MS n spectra. At a false discovery rate of less than 0.01, HiTMS correctly assigned 85% of the structures in a library of 133 manually annotated Francisella tularensis subspecies novicida lipid A structures. Additionally, HiTMS correctly assigned 85% of the structures in a smaller library of lipid A species from Yersinia pestis demonstrating that it may be used across species.

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Detergent-accelerated hydrolysis of bacterial endotoxins and determination of the anomeric configuration of the glycosyl phosphate present in the “Isolated lipid A” fragment of the Bordetella pertussis endotoxin

Cited by 221 publications

References 27 publications

Function of Escherichia coli MsbA, an Essential ABC Family Transporter, in Lipid A and Phospholipid Biosynthesis

Function of Escherichia coli MsbA, an Essential ABC Family Transporter, in Lipid A and Phospholipid Biosynthesis

Deciphering tissue-induced Klebsiella pneumoniae lipid A structure

Automated Lipid A Structure Assignment from Hierarchical Tandem Mass Spectrometry Data

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