Proteomic Analysis of Helicobacter pylori Whole Cell Proteins using the Narrow Range IPG Strips.
10.4167/jbv.2007.37.4.203
- Author:
Jeong Won PARK
1
;
Seung Gyu LEE
;
Jae Young SONG
;
Jin Su JUN
;
Jung Soo JOO
;
Hee Shang YOUN
;
Ji Hyun SEO
;
Hyung Lyun KANG
;
Seung Chul BAIK
;
Woo Kon LEE
;
Myung Je CHO
;
Kwang Ho RHEE
Author Information
1. Department of Microbiology, Gyeongsang National University School of Medicine, Jinju, Gyeong-Nam 660-751, Republic of Korea. khrhee@gaechuk.gsnu.ac.kr
- Publication Type:Original Article
- Keywords:
Narrow range IPG strip;
Helicobacter pylori;
Proteomics
- MeSH:
Chemotaxis;
Electrophoresis, Polyacrylamide Gel;
Helicobacter pylori*;
Helicobacter*;
Hydrogen-Ion Concentration;
Hydrogenase;
Peptide Mapping;
Proteome;
Proteomics;
Ribosomal Protein S6;
Silver Staining
- From:Journal of Bacteriology and Virology
2007;37(4):203-212
- CountryRepublic of Korea
- Language:English
-
Abstract:
It has been reported that most of Helicobacter pylori proteome components appear so crowded in the region of pH 4.5~8.0 that a lot of them were inseparable in 2-DE using the broad range IPG strip. Therefore, inseparable protein spots in 2-DE profiles have to be apart from each other for improving the protein identification. Here, we attempt to examine the usability of the narrow range IPG strips for separating close spots in the broad range IPG strip at proteomic analysis of H. pylori. The whole cell proteins of H. pylori strain 26695 were separated by narrow range IPG strips (pI 3.9~5.1, 4.7~5.9, 5.5~6.7, and 6.3~8.3, respectively), followed by SDS-PAGE, and visualized by silver staining, showing that the distances between spots were widened and the total number of detectable spots was increased. Resolved protein spots were identified by the peptide fingerprinting using MALDI-TOF-MS. As a result, 87 expressed proteins were identified by the peptide fingerprinting. Of them, 23 proteins, including hydrogenase expression/formation protein, purine-binding chemotaxis protein, and ribosomal protein S6, have not been reported in the previous proteome studies of H. pylori. Thus, these results demonstrate that the high complexity proteome components could be effectively separated using the narrow range IPG strips, which might be helpful to strengthen the contents of the master protein map of the H. pylori reference strain.
