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World J Gastroenterol. Nov 21, 2005; 11(43): 6880-6883
Published online Nov 21, 2005. doi: 10.3748/wjg.v11.i43.6880
Immunoproteomics of membrane proteins of Shigella flexneri 2a 2457T
Tian-Yi Ying, Jun-Jun Wang, Heng-Liang Wang, Er-Ling Feng, Kai-Hua Wei, Liu-Yu Huang, Pei-Tang Huang, Cui-Fen Huang
Tian-Yi Ying, Jun-Jun Wang, Heng-Liang Wang, Er-Ling Feng, Kai-Hua Wei, Liu-Yu Huang, Pei-Tang Huang, Cui-Fen Huang, Beijing Institute of Biotechnology, State Key Lab of Pathogen and Biosecurity, Beijing 100071, China
Tian-Yi Ying, Beijing Institute of Pharmaceutical Chemistry, Beijing 102205, China
Author contributions: All authors contributed equally to the work.
Supported by the Capital “248” Key Innovation Project, No. H010210360119, State Basic Research Development Program of China No. 973 Program, G1999054103 and 2005CB22904 and National Natural Science Foundation of China No. 30470101
Correspondence to: Dr Heng-Liang Wang, Institute of Biotechnology, State Key Lab of Pathogen and Biosecurity, 20 Dongdajie Street, Fengtai District, Beijing 100071, China. wanghl@nic.bmi.ac.cn
Telephone: +86-10-66948836
Received: April 12, 2005
Revised: April 28, 2005
Accepted: April 30, 2005
Published online: November 21, 2005
Abstract

AIM: 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.

CONCLUSION: Membrane proteins of S. flexneri 2a 2457T were successfully observed by 2-DE. Several known and novel antigens were identified by mass spectrum.

Keywords: Shigella flexneri 2a 2457T, Immunoproteomics, Membrane proteins