Immunoproteome analysis of soluble and membrane proteins of Shigella flexneri 2457T.
OBJECTIVE
To profile the immunogenic proteins of Shigella flexneri (S. flexneri) expressed during human infection using a proteomic approach.


METHODS
Soluble and membrane protein extractions of S. flexneri 2457T were separated by two-dimensional gel electrophoresis (2-DE). Proteins were transferred to PVDF membrane and immunoblotted with sera from shigellosis patients. Reactive protein spots were matched to Coomassie stained gels run in parallel, cut out and trypsin digested. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) was used to determine the peptide mass fingerprints, which were searched in the MASCOT database to identify the protein.


RESULTS
A total of 8 immunoreactive proteins were successfully identified from the Coomassie stained gels in three repeats. Six of these proteins have not previously been reported as immunogenic in S. flexneri. These proteins could be potential candidates for vaccine or attenuation studies.


CONCLUSIONS
Soluble and membrane proteins of S. flexneri 2457T have been screened by 2-DE and immunoblotting with sera from shigellosis patients. Eight proteins are identified as immunogenic.
Jennison
Amy V
AV
School of Biochemistry and Molecular Biology, Faculty of Science, The Australian National University, Canberra, ACT 0200, Australia.
Raqib
Rubhana
R
Verma
Naresh K
NK
D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D001428:Bacterial Vaccines / Q000276:immunology; D004269:DNA, Bacterial / Q000235:genetics; D030541:Databases, Genetic; D004405:Dysentery, Bacillary / Q000097:blood / Q000517:prevention & control; D015180:Electrophoresis, Gel, Two-Dimensional; D015151:Immunoblotting; D007125:Immunogenetics / Q000379:methods; D008565:Membrane Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D010449:Peptide Mapping; D020543:Proteome / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D012763:Shigella flexneri / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D012995:Solubility
D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D004269:DNA, Bacterial; D008565:Membrane Proteins; D020543:Proteome