Immunoproteomics of membrane proteins of Shigella flexneri 2a 2457T.
OBJECTIVE
To screen the immunogenic membrane proteins of Shigella flexneri 2a 2457T.


METHODS
The routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western blotting were combined to screen immunogenic proteins of S. flexneri 2a 2457T. Serum was gained from rabbits immunized with the same bacteria. Immunogenic spots were cut out from the polyacrylamide gel and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) was performed to determine the molecular weight of peptides. Electrospray ionization (ESI-MS/MS) was performed to determine the sequences of the interesting peptides.


RESULTS
A total of 20 spots were successfully identified from Coomassie brilliant blue stained gels representing 13 protein entries, 5 known antigens and 8 novel antigens. A hypothetical protein (YaeT) was detected, which might be a candidate target of vaccine.


CONCLUSIONS
Membrane proteins of S. flexneri 2a 2457T were successfully observed by 2-DE. Several known and novel antigens were identified by mass spectrum.
Ying
Tian-Yi
TY
Beijing Institute of Biotechnology, State Key Lab of Pathogen and Biosecurity, Beijing 100071, China.
Wang
Jun-Jun
JJ
Wang
Heng-Liang
HL
Feng
Er-Ling
EL
Wei
Kai-Hua
KH
Huang
Liu-Yu
LY
Huang
Pei-Tang
PT
Huang
Cui-Fen
CF
D000818:Animals; D000942:Antigens, Bacterial / Q000737:chemistry / Q000276:immunology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000276:immunology*; D002648:Child; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000276:immunology; D040901:Proteomics / Q000379:methods*; D011817:Rabbits; D012763:Shigella flexneri / Q000276:immunology*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D008565:Membrane Proteins