Cholera is a life-threatening diarrhoeal disease caused by ingestion of Vibrio cholerae. There are at least 200 serogroups of V. cholerae but only two of them are causing epidemics – O1 and O139 serogroups. Fragmentation analysis of O-antigen, also known as O-specific polysaccharide (OSP), from lipopolysaccharide (LPS) is important to obtain new information about its structure, such as fragmentation patterns and fragment structures. In the present study, OSP and core (OSPc) structure from V. cholerae O139 was studied using matrix-assisted laser desorption ionization (MALDI)-time of flight (TOF) and direct injection electrospray ionization (ESI)-MS methods. MALDI-TOF analysis was performed in positive-ion reflectron mode, while ESI-MS was performed in negative ionization mode. ESI-MS analysis was followed by ESI-MS/MS analysis. Using this analytical approach, we managed to obtain two possible fragmentation pathways of OSP from V. cholerae O139. Mutual sign of these two pathways is shortening the length of the oligosaccharide by neutral loss of monosaccharide residues. Additionally, liquid chromatography-MS analysis was performed to separate depicted molecular forms of OSPc.
DeenJMengelMAClemensJD. Epidemiology of cholera. Vaccine. 2020; 38: A31–A40. doi:10.1016/j.vaccine.2019.07.078
2.
ConnorBADawoodRRiddleMS,et al.Cholera in travellers: a systematic review. J Travel Med2019; 26. doi: 10.1093/jtm/taz085
3.
RamamurthyTMutrejaAWeillFX,et al.Revisiting the global epidemiology of cholera in conjuction with the genomics of Vibrio cholerae. Front Public Health2019; 7: 03.
4.
BhandareSColomJBaigA, et al.Reviving phage therapy for the treatment of cholera. J Infect Dis2019; 219: 786–794.
5.
LopezALGonzalesMLAldabaJG,et al.Killed oral cholera vaccines: history, development and implementation challenges. Ther Adv Vaccines2014; 2: 123–136.
6.
GrandjeanCBoutonnierADassyB,et al.Investigation towards bivalent chemically defined glycoconjugate immunogens prepared from acid-detoxified lipopolysaccharide of Vibrio cholerae O1, serotype Inaba. Glycoconj J2009; 26: 41–55.
7.
KayECuccuiJWrenBW. Recent advances in the production of recombinant glycoconjugate vaccines. npj Vaccines2019; 4. doi: 10.1038/s41541-019-0110-z
8.
LuXPfisterHBSolimanSE,et al.O-Specific polysaccharide of Vibrio cholerae O139: improved synthesis and conjugation to BSA by squaric acid chemistry. Eur J Org Chem2018: 2944–2957. doi: 10.1002/ejoc.201800429
9.
SolimanSEKováčP. Total synthesis of the complete protective antigen of Vibrio cholerae O139. Chem Int Ed2016; 55: 12850–12853.
10.
KnirelYAWidmalmGSenchenkovaSN,et al.Structural studies on the short-chain lipopolysaccharide of Vibrio cholerae 0139 Bengal. Eur J Biochem1997; 247: 402–410.
11.
CoxADBrissonJRVarmaV,et al.Structural analysis of the lipopolysaccharide from Vibrio cholerae O139. Carbohydr Res1996; 290: 43–58.
12.
CoxADPerryMB. Structural analysis of the O-antigen-core region of the lipopolysaccharide from Vibrio cholerae O139. Carbohydr Res1996; 290: 59–65.
13.
XuPKorcováJBaráthP, et al.Isolation, purification, characterization and direct conjugation of the lipid A-free lipopolysaccharide of Vibrio cholerae O139. Chemistry2019; 25: 12946–12956.
14.
ChmelíkJŘehulkaPKováčikV,et al.Negative matrix-assisted laser desorption/ionization time-of-flight/time-of-flight tandem mass spectrometry fragmentation of synthetic analogs of the O-specific polysaccharide of Vibrio cholerae O:1 in the presence of anionic dopants. Eur J Mass Spectrom (Chichester)2007; 13: 347–353.
15.
KováčikVBekešováSTvaroškaI,et al.Positive electrospray ion trap multistage mass spectrometric fragmentation of synthetic analogs of saccharide part of lipopolysaccharides of Vibrio cholerae O:1. J Am Soc Mass Spectrom2006; 17: 749–756.
16.
BekesováSKovácikVChmelíkJ,et al.Negative electrospray, ion trap multistage mass spectrometry of synthetic fragments of the O-PS of Vibrio cholerae O:1. Eur J Mass Spectrom (Chichester)2006; 12: 43–50.
17.
PhillipsNJJohnCMJarvisGA. Analysis of bacterial lipooligosaccharides by MALDI-TOF MS with traveling wave ion mobility. J Am Soc Mass Spectrom2016; 27: 1263–1276.
18.
ChoiYRKimKSBanduR, et al.Liquid chromatography/electrospray ionization tandem mass spectrometry-based structural analysis of deacylated lipooligosaccharides from escherichia coli. Bull Korean Chem Soc2020; 41: 454–459.
19.
O’BrienJPNeedhamBDBrownDB,et al.Top-down strategies for the structural elucidation of intact gram-negative bacterial endotoxins. Chem. Sci2014; 5: 4291–4301.
20.
PostDMYuLKrasityBC, et al.O-antigen and core carbohydrate of Vibrio fischeri lipopolysaccharide: composition and analysis of their role in Euprymna scolopes light organ colonization. J Biol Chem2012; 287: 8515–8530.
21.
DomonBCostelloCE. A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MSSpectra of glycoconjugates. Glycoconjugate J1988; 5: 397–409.
22.
LuXKováčP. Chemical synthesis of the galacturonic acid containing pentasaccharide antigen of the O-specific polysaccharide of Vibrio cholerae O139 and its five fragments. J Org Chem2016; 81: 6374–6394.
