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Kim, Lee, Joo, Kwon, Bea, Song, Kang, Lee, Cho, Rhee, Youn, and Baik: Proteomic analysis of Helicobacter pylori J99 Outer Membrane Protein by Tandem Mass Spectrometry
Original Article

J Bacteriol Virol 2008;38(2):53-60.

Published online: 18 February 2008

DOI: https://doi.org/10.4167/jbv.2008.38.2.53

Proteomic analysis of Helicobacter pylori J99 Outer Membrane Protein by Tandem Mass Spectrometry

Kyung-Mi Kim1, Seung-Gyu Lee1, Jung-Soo Joo1, Young-Chul Kwon1, Dong-Won Bea3, Jea-Young Song1, 4, Hyung-Lyun Kang1, 4, Woo-Kon Lee1, 4, Myung-Je Cho1, 4, Kwang-Ho Rhee1, 4, Hee-Shang Youn2, Seung-Chul Baik1, 4, ∗

1Department of Microbiology, Gyeongsang National University College of Medicine, Jinju, Gyeongsangnam-do, Republic of Korea

2Department of Pediatrics, Gyeongsang National University College of Medicine, Jinju, Gyeongsangnam-do, Republic of Korea

3Central Instrument Facility, Gyeongsang National University, Jinju, Gyeongsangnam-do, Republic of Korea

4Research Institute of Life Science, Gyeongsang National University, Jinju, Gyeongsangnam-do, Republic of Korea

∗Corresponding author: Seung-Chul Baik. Department of Microbiology, Gyeongsang National University College of Medicine, Chiram-dong 90, Jinju, Gyeongsangnam-do, 660-751, Republic of Korea. Phone: +82-55-751-8780, Fax: +82-55-759-1588, e-mail: scbaik@gaechuk.gsnu.ac.kr

∗∗ This work was supported by a grant R01-2003-000-10533-0 from KOSEF.

Received 3 May 2008       Revised 17 May 2008       Accepted 23 May 2008

Copyright © 2008 Journal of Bacteriology and Virology

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

The protein identity of sarcosine-insoluble outer membrane proteins (OMPs) of Helicobacter pylori J99 was determined with the basic study of understanding the function of proteins. A sarcosine-insoluble OMPs was resolved by two-dimensional electrophoresis with immobilized pH gradient strips. The most abundant proteins were shown in the alkaline pI regions (6.0∼11.0) with molecular masses of 10 to 100 kDa. We have performed an extensive proteome analysis by quadrupole time of flight (Q-TOF) mass spectrometry (MS). Here, of 50 spots processed, 42 spots were identified, which represented 16 genes and we newly detected 8 kinds of proteins (JHP0119, JHP0388, JHP1046, JHP1405, JHP0073, JHP0551, JHP1382, JHP0552) from the sarcosin-insoluble fraction of H. pylori J99. Those may be used to elucidate the characterization of the OMPs of H. pylori J99, which will help identify new potential target proteins for vaccine development and drug therapy.

Keywords: Helicobacter pylori, Outer membrane protein, Q-TOF MS

References

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Figure 1.
2-DE of sarcosine-insoluble outer membrane fraction of H. pylori J99 with an IPG strip, pH 6.0 to11.0. Spots that were analyzed by Q-TOF MS are numbered.
jbv-38-53f1.tif
Table 1.
List of identified proteins in the sarcosine-insoluble outer membrane fraction of H. pylori J99
Spot no Protein identification Accession noa No. of peptide matches Scoreb and seq. C (%)c Amino acid sequence TpId TMrd JHP noa
1 Putative outer membrane protein gi|15611491 1 37, 5.4 FQFLWNLGGR 9.39 20895 JHP0424
2 Hypothetical protein gi|15612165 1 36, 4.7 FSYEDSLLK 9.46 20838 JHP1100
3 Hypothetical protein gi|15612165 1 38, 5.9 SFIDGDLDIQK 9.46 20838 JHP1100
4 Hypothetical protein gi|15612165 1 89, 15.1 GQVITLIGQNEVPYLILETDCQVGDIAK 9.46 20838 JHP1100
5 Hypothetical protein gi|15611189 1 61, 3.9 GDLSAFGAFFK 9.93 32642 JHP0119
6 Hypothetical protein gi|15611189 1 47, 4.6 K.GDLSAFGAFFK.G 9.93 32642 JHP0119
7 Hypothetical protein gi|15611189 1 30, 4.6 K.GDLSAFGAFFK.G 9.93 32642 JHP0119
8 Hypothetical protein gi|15611189 1 20, 4.6 K.GDLSAFGAFFK.G 9.93 32642 JHP0119
9 Hypothetical protein gi|15611189 1 35, 4.6 K.GDLSAFGAFFK.G 9.93 32642 JHP0119
10 Gamma-glutamyltranspeptidase (ggt) gi|15612111 1 32, 2.1 VFLVVGSPGGSR 9.72 61089 JHP1046
11 Hypothetical protein gi|15611189 3 135, 14.4 K.GDLSAFGAFFK.G
K.VSFVVNDR.E
K.DLGTELSLPLFNWLYK.G		 9.93 32642 JHP0119
12 Hypothetical protein gi|15611189 2 79, 10.9 GDLSAFGAFFK
GSDFGALHEQFGDMYDGYIK		 9.93 32642 JHP0119
13 Hypothetical protein gi|15611189 1 23, 7.0 GSDFGALHEQFGDMYDGYIK 9.93 32642 JHP0119
14 Hypothetical protein gi|15611189 1 41, 3.9 GDLSAFGAFFK 9.93 32642 JHP0119
15 Outer membrane protein-adhesin (babA) gi|15611900 3 132, 5.2 YSTLNTLIK
LSADPSAINAVR
GIQDLSDRYESLNNLLNR		 9.16 80607 JHP0833
16 Outer membrane protein-adhesin (babA) gi|15611900 2 88, 2.8 LSADPSAINAVR
YSTLNTLIK		 9.16 80607 JHP0833
17 Putative outer membrane protein gi|15612427 2 47, 9.7 VYAFQISYLR
FPPYAGPGFEVGYK		 9.90 28238 JHP1362
18 Outer membrane protein-porin (hopE) gi|15611712 2 63, 13.3 K.YANGALNGFGLNVGYK.K
K.FLSAGPNATNLYYHLK.R		 9.11 29551 JHP0645
19 Outer membrane protein-porin (hopE) gi|15611712 1 30, 5.9 FLSAGPNATNLYYHLK 9.11 29551 JHP0645
20 Outer membrane protein-porin (hopE) gi|15611712 1 40, 5.9 FLSAGPNATNLYYHLK 9.11 29551 JHP0645
21 Putative outer membrane protein gi15611144 2 72, 9.8 R.GVDGSVDVFYK.R
K.LPLFTNQFYK.E		 9.05 28351 JHP0073
22 Outer membrane protein/porin (hopB) gi|15611916 3 100, 12 K.FQFLFDVGLR.M
K.SHNQHSIEIGVQIPTIYNTYYK.A
K.ANPWLGNFAAGNSSQVNAFNGFITK.I		 9.33 56753 JHP0849
23 Outer membrane protein/porin (hopB) gi|15611916 1 52, 2.3 K.FQFLFDVGLR.M 9.33 56753 JHP0849
24 Outer membrane protein/porin (hopB) gi|15611916 2 76, 7.4 K.FQFLFDVGLR.M
K.ANPWLGNFAAGNSSQVNAFNGFITK.I		 9.33 56753 JHP0849
25 Putative outer membrane protein gi|15612459 1 45, 2.6 IPTLPNYFFK 9.75 42902 JHP1394
26 Putative outer membrane protein gi|15612459 2 78, 9.3 IPTLPNYFFK
QGPLENGNPTTITGAETNFSLTQTLR		 9.75 42902 JHP1394
27 Putative outer membrane protein gi|15612459 1 26, 3.1 K.IPTLPNYFFK.G 9.75 42902 JHP1394
28 Putative outer membrane protein gi|15612459 1 54, 3.1 K.IPTLPNYFFK.G 9.75 42902 JHP1394
29 Hypothetical protein gi|15611619 2 87, 5.7 K.TTIDAPNLQLR.E
R.LTLEYLTNLSVK.N		 9.39 54624 JHP0552
30 Putative gi|15612447 2 73, 5.0 R.NTLSSIIIVEQK.S
K.SLLSSVELAK.E		 9.51 56924 JHP1382
31 Putative iron regulated outer membrane protein (frpB_3) gi|15612470 3 100, 5.6 R.VESTAFLGVR.G
R.YDIYTLLDK.N
R.THVTSGFSPSATVLYNPIESIGLK.V		 9.16 97572 JHP1405
32 Putative iron regulated outer membrane protein (frpB_3) gi|15612470 2 77, 2.8 R.VESTAFLGVR.G
R.IFLINSGVNVK.V		 9.16 97572 JHP1405
33 Putative iron regulated outer membrane protein (frpB_3) gi 15612470 2 71, 2.6 R.VESTAFLGVR.G
R.YDIYTLLDK.N		 9.16 97572 JHP1405
34 Putative iron regulated outer membrane protein (frpB_3) gi 15612470 3 108, 4.1 R.VESTAFLGVR.G
R.IFLINSGVNVK.V
R.YDIYTLLDK.N		 9.16 97572 JHP1405
35 Putative iron regulated outer membrane protein (frpB_3) gi 15612470 3 108, 4.1 R.VESTAFLGVR.G
R.YDIYTLLDK.N
R.IFLINSGVNVK.V		 9.16 97572 JHP1405
36 Putative outer membrane function gi|15612168 1 47, 2.3 IPTINTNYYSYLGTK 9.51 69547 JHP1103
37 Hypothetical protein gi 15611619 2 42, 5.2 K.AALGLYELLK.G
K.TTIDAPNLQLR.E		 9.39 54624 JHP0552
38 Hypothetical protein gi|15611619 4 121, 11.5 K.AALGLYELLK.G
K.TTIDAPNLQLR.E
R.LTLEYLTNLSVK.N
K.ASLDAANLSFANIK.R		 9.39 54624 JHP0552
39 Hypothetical protein gi 15611619 2 47, 5.2 K.AALGLYELLK.G
K.TTIDAPNLQLR.E		 9.39 54624 JHP0552
40 Outer membrane protein/porin (hopC) gi|15611915 1 20, 2.5 R.ATNILNGFYTK.V 9.36 56404 JHP0848
41 Outer membrane protein/porin (hopB)   2 104, 5.3 K.FQFLFDVGLR.M
R.STQLLNNTTNTLAK.V		 9.33 56753 JHP0849
42 Putative outer membrane protein gi 15612326 1 54, 1.9 YNQLQTVAQELGK 6.52 75659 JHP1261

a accession number and JHP number are obtained from NCBI database;

b score, probability score in Mascot program;

c seq. C (%), percentage of sequence coverage;

d TpI and T Mr, Theoretical pI and Mr are from www.tigr.org.

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