Article (Scientific journals)
Use of Capillary Zone Electrophoresis Coupled to Electrospray Mass Spectrometry for the Detection and Absolute Quantitation of Peptidoglycan-Derived Peptides in Bacterial Cytoplasmic Extracts
Delvaux, Cédric; Dauvin, Marjorie; Boulanger, Madeleine et al.
2021In Analytical Chemistry, 93 (4), p. 2342-2350
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Abstract :
[en] Peptidoglycan (PGN) is an essential structure found in the bacterial cell wall. During the bacterial life cycle, PGN continuously undergoes biosynthesis and degradation to ensure bacterial growth and division. The resulting PGN fragments (muropeptides and peptides), which are generated by the bacterial autolytic system, are usually transported into the cytoplasm to be recycled. On the other hand, PGN fragments can act as messenger molecules involved in the bacterial cell wall stress response as in the case of β-lactamase induction in the presence of β-lactam antibiotic or in triggering mammalian innate immune response. During their cellular life, bacteria modulate their PGN degradation by their autolytic system or their recognition by the mammalian innate immune system by chemically modifying their PGN. Among these modifications, the amidation of the ε-carboxyl group of meso-diaminopimelic acid present in the PGN peptide chain is frequently observed. Currently, the detection and quantitation of PGN-derived peptides is still challenging because of the difficulty in separating these highly hydrophilic molecules by RP-HPLC as these compounds are eluted closely after the column void volume or coeluted in many cases. Here, we report the use of capillary zone electrophoresis coupled via an electrospray-based CE–MS interface to high-resolution mass spectrometry for the quantitation of three PGN peptides of interest and their amidated derivatives in bacterial cytoplasmic extracts. The absolute quantitation of the tripeptide based on the [13C,15N] isotopically labeled standard was also performed in crude cytoplasmic extracts of bacteria grown in the presence or absence of a β-lactam antibiotic (cephalosporin C). Despite the high complexity of the samples, the repeatability of the CZE–MS quantitation results was excellent, with relative standard deviations close to 1%. The global reproducibility of the method including biological handling was better than 20%
Research center :
laboratoire de spectrométrie de masse (MSLab)
CIP - Centre d'Ingénierie des Protéines - ULiège
Université Paris-Saclay, CEA, CNRS, Institute for Integrative Biology of the Cell (I2BC), Gif-sur-Yvette 91198, France
Disciplines :
Chemistry
Author, co-author :
Delvaux, Cédric  ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
Dauvin, Marjorie ;  Université de Liège - ULiège > Département des sciences de la vie > Centre d'ingénierie des protéines > Ph.D
Boulanger, Madeleine ;  Université de Liège - ULiège > Département des sciences de la vie > Centre d'ingénierie des protéines > Ph.D
Quinton, Loïc  ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie biologique
Mengin-Lecreulx, Dominique;  Université Paris-Saclay > CEA, CNRS > Institute for Integrative Biology of the Cell (I2BC)
Joris, Bernard ;  Université de Liège - ULiège > Département des sciences de la vie > Centre d'ingénierie des protéines
De Pauw, Edwin  ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
Far, Johann  ;  Université de Liège - ULiège > Département de chimie (sciences) > Chimie analytique inorganique
Language :
English
Title :
Use of Capillary Zone Electrophoresis Coupled to Electrospray Mass Spectrometry for the Detection and Absolute Quantitation of Peptidoglycan-Derived Peptides in Bacterial Cytoplasmic Extracts
Publication date :
20 January 2021
Journal title :
Analytical Chemistry
ISSN :
0003-2700
eISSN :
1520-6882
Publisher :
American Chemical Society, United States - District of Columbia
Volume :
93
Issue :
4
Pages :
2342-2350
Peer reviewed :
Peer Reviewed verified by ORBi
Name of the research project :
ARC NetRBI
Funders :
ULiège - Université de Liège [BE]
Available on ORBi :
since 07 July 2021

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