Proteomic characterisation of the Chlamydia abortus outer membrane complex (COMC) using combined rapid monolithic column liquid chromatography and fast MS/MS scanning

Autor: Lisa Imrie, Morag Livingstone, Neil F. Inglis, Kevin Aitchison, David Longbottom, Nick Wheelhouse, Erin D. T. Manson
Rok vydání: 2019
Předmět:
Proteome
Cell Membranes
Pathology and Laboratory Medicine
Physical Chemistry
Chaperonin
Chlamydia Infection
Pregnancy
Tandem Mass Spectrometry
Medicine and Health Sciences
Chlamydia
Peptide sequence
0303 health sciences
Multidisciplinary
Chemistry
General Medicine
Bacterial Pathogens
Molecular Mass
Infectious Diseases
Chlamydia Trachomatis
Medical Microbiology
Physical Sciences
Medicine
Female
Cellular Structures and Organelles
Pathogens
Bacterial outer membrane
General Agricultural and Biological Sciences
Bacterial Outer Membrane Proteins
Research Article
Monolithic HPLC column
Science
Sexually Transmitted Diseases
Mass spectrometry
Microbiology
General Biochemistry
Genetics and Molecular Biology
03 medical and health sciences
Antigen
Animals
Integral Membrane Proteins
Fragmentation (cell biology)
Microbial Pathogens
Gram Negative Bacteria
030304 developmental biology
Chromatography
Sheep
Bacteria
030306 microbiology
Organisms
Biology and Life Sciences
Membrane Proteins
Bacteriology
Cell Biology
Chlamydia Infections
Outer Membrane Proteins
Membrane protein
Chemical Properties
Chromatography
Liquid
Zdroj: PLoS ONE
PLoS ONE, Vol 14, Iss 10, p e0224070 (2019)
ISSN: 1932-6203
Popis: Data are presented on the identification and partial characterisation of proteins comprising the chlamydial outer membrane complex (COMC) fraction of Chlamydia abortus (C. abortus)-the aetiological agent of ovine enzootic abortion. Inoculation with the COMC fraction is known to be highly effective in protecting sheep against experimental challenge and its constituent proteins are therefore of interest as potential vaccine candidates. Sodium N-lauroylsarcosine (sarkosyl) insoluble COMC proteins resolved by SDS-PAGE were interrogated by mass spectrometry using combined rapid monolithic column liquid chromatography and fast MS/MS scanning. Downstream database mining of processed tandem MS data revealed the presence of 67 proteins in total, including putative membrane associated proteins (n = 36), such as porins, polymorphic membrane proteins (Pmps), chaperonins and hypothetical membrane proteins, in addition to others (n = 22) that appear more likely to have originated from other subcellular compartments. Electrophoretic mobility data combined with detailed amino acid sequence information derived from secondary fragmentation spectra for 8 Pmps enabled peptides originating from protein cleavage fragments to be mapped to corresponding regions of parent precursor molecules yielding preliminary evidence in support of endogenous post-translational processing of outer membrane proteins in C. abortus. The data presented here will facilitate a deeper understanding of the pathogenesis of C. abortus infection and represent an important step towards the elucidation of the mechanisms of immunoprotection against C. abortus infection and the identification of potential target vaccine candidate antigens.
Databáze: OpenAIRE
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