Dissecting Lipopolysaccharide Composition and Structure by GC-MS and MALDI Spectrometry

García-Vello, Pilar; Speciale, Immacolata; Lorenzo, Flaviana Di; Molinaro, Antonio; Castro, Cristina De

doi:10.1007/978-1-0716-2581-1_12

Cited by 11 publications

(9 citation statements)

References 49 publications

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“…Such preparations are analyzed by means of GLC-MS and reveal the presence and type of monosaccharides and fatty acids. 57 In the next step, monosaccharides and fatty acids are quantitatively analyzed using internal and external standards. As an alternative, monosaccharides are measured in GLC or GC-MS as alditol acetates obtained after cleavage of the glycosidic linkages, reduction with sodium borohydride, and peracetylation.…”

Section: Compositional Analysesmentioning

confidence: 99%

“…Typically, when data attained from these MS approaches are merged with data from monosaccharide and fatty acid compositional analysis, it is possible to define the complete structure of a lipid A. 57 However, to gain detailed structural information, such as the location of a specific secondary acyl substituent or even of a substituent decorating the lipid A sugar backbone, the employment of MS 2 approaches is mandatory. Notably, the development of new methods for sample and matrix preparation allows to obtain high-quality MALDI mass spectra of intact LOS, which provide a clear picture of the entire macromolecule without prior chemical degradations.…”

Section: Structural Analysis Of Lpsmentioning

confidence: 99%

“…To obtain an overview of the composition of the tested LPS sample, it is feasible to start with the analysis of acetylated O -methyl glycosides that are formed after glycosidic cleavage with hydrochloric acid in anhydrous methanol and peracetylation with acetic anhydride in pyridine. Such preparations are analyzed by means of GLC-MS and reveal the presence and type of monosaccharides and fatty acids . In the next step, monosaccharides and fatty acids are quantitatively analyzed using internal and external standards.…”

Section: Chemistry Of the Lps: The Structural Characterizationmentioning

confidence: 99%

“…MS has played a central role in characterizing LPS from different organisms with MS-based approaches still as the most frequently employed to characterize lipid A structures. , Several ionization techniques are commonly used to analyze LPS and its structural components: electron ionization (EI), chemical ionization (CI), electrospray ionization (ESI), and matrix-assisted laser desorption/ionization (MALDI). Typically, when data attained from these MS approaches are merged with data from monosaccharide and fatty acid compositional analysis, it is possible to define the complete structure of a lipid A . However, to gain detailed structural information, such as the location of a specific secondary acyl substituent or even of a substituent decorating the lipid A sugar backbone, the employment of MS 2 approaches is mandatory.…”

Section: Chemistry Of the Lps: The Structural Characterizationmentioning

confidence: 99%

See 3 more Smart Citations

A Journey from Structure to Function of Bacterial Lipopolysaccharides

et al. 2021

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Lipopolysaccharide (LPS) is a crucial constituent of the outer membrane of most Gram-negative bacteria, playing a fundamental role in the protection of bacteria from environmental stress factors, in drug resistance, in pathogenesis, and in symbiosis. During the last decades, LPS has been thoroughly dissected, and massive information on this fascinating biomolecule is now available. In this Review, we will give the reader a third millennium update of the current knowledge of LPS with key information on the inherent peculiar carbohydrate chemistry due to often puzzling sugar residues that are uniquely found on it. Then, we will drive the reader through the complex and multifarious immunological outcomes that any given LPS can raise, which is strictly dependent on its chemical structure. Further, we will argue about issues that still remain unresolved and that would represent the immediate future of LPS research. It is critical to address these points to complete our notions on LPS chemistry, functions, and roles, in turn leading to innovative ways to manipulate the processes involving such a still controversial and intriguing biomolecule.

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Section: Compositional Analysesmentioning

confidence: 99%

Section: Structural Analysis Of Lpsmentioning

confidence: 99%

Section: Chemistry Of the Lps: The Structural Characterizationmentioning

confidence: 99%

Section: Chemistry Of the Lps: The Structural Characterizationmentioning

confidence: 99%

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A Journey from Structure to Function of Bacterial Lipopolysaccharides

et al. 2021

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“…In addition, an aliquot of the isolated lipid A fractions underwent methanolysis (1.25 M HCl/CH 3 OH, 80 °C, 16 h), followed by acetylation (80 °C, 20 min) and GC-MS analysis. [45,46] The D-configuration of the mannose and the glucosamine constituting the lipid A saccharide backbone was defined by GC-MS analysis of the acetylated O-(+)-2-octyl glycoside derivatives and comparison with authentic standards, as previously described. [47] The temperature program for GC-MS analysis used to analyze the sugar content was as follows: 140 °C for 3 min and then 140 °C !240 °C at 3 °C/min.…”

Section: Isolation and Analysis Of The Lipid A Fractionsmentioning

confidence: 99%

Pushing the Boundaries of Structural Heterogeneity with the Lipid A of Marine Bacteria Cellulophaga

et al. 2023

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Marine bacteria, which are often described as chemical gold, are considered an exceptional source of new therapeutics. Considerable research interest has been given to lipopolysaccharides (LPSs), the main components of the Gram-negative outer membrane. LPS and its lipid A portion from marine bacteria are known to exhibit a tricky chemistry that has been often associated with intriguing properties such as behaving as immune adjuvants or anti-sepsis molecules. In this scenario, we report the structural determination of the lipid A from three marine bacteria within the Cellulophaga genus, which showed to produce an extremely heterogenous blend of tetra-to hexaacylated lipid A species, mostly carrying one phosphate and one D-mannose on the glucosamine disaccharide backbone. The ability of the three LPSs in activating TLR4 signaling revealed a weaker immunopotential by C. baltica NNO 15840 T and C. tyrosinoxydans EM41 T , while C. algicola ACAM 630 T behaved as a more potent TLR4 activator.

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Deciphering the Chemical Language of the Immunomodulatory Properties of Veillonella parvula Lipopolysaccharide

Pither,

Andretta,

Rocca

et al. 2024

Angewandte Chemie

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Veillonella parvula, prototypical member of the oral and gut microbiota, is at times commensal yet also potentially pathogenic. The definition of the molecular basis tailoring this contrasting behavior is key for broadening our understanding of the microbiota driven pathogenic and/or tolerogenic mechanisms that take place within our body. In this study, we focused on the chemistry of the main constituent of the outer membrane of V. parvula, the lipopolysaccharide (LPS). LPS molecules indeed elicit pro‐inflammatory and immunomodulatory responses depending on their chemical structures. Here we report about the structural elucidation of the LPS from two strains of V. parvula and show important and unprecedented differences in both the lipid and carbohydrate moieties, including the identification of a novel galactofuranose and mannitol‐containing O‐antigen repeating unit for one of the two strains. Furthermore, by harnessing computational studies, in vitro human cell models, as well as lectin binding solid phase assays, we discovered that the two chemically diverse LPS immunologically behave differently and have attempted to identify the molecular determinant(s) governing this phenomenon. Whether a pro‐inflammatory potential has been evidenced for the lipid A moiety, by contrast a plausible “immune modulating” action has been proposed for the peculiar O‐antigen portion.

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Dissecting Lipopolysaccharide Composition and Structure by GC-MS and MALDI Spectrometry

Cited by 11 publications

References 49 publications

A Journey from Structure to Function of Bacterial Lipopolysaccharides

A Journey from Structure to Function of Bacterial Lipopolysaccharides

Pushing the Boundaries of Structural Heterogeneity with the Lipid A of Marine Bacteria Cellulophaga

Deciphering the Chemical Language of the Immunomodulatory Properties of Veillonella parvula Lipopolysaccharide

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