Proteome Analysis of Alkylhydroxide Peroxidase-Deficient Isogenic Mutant of Helicobacter pylori 26695

Woo-Kon Lee; Seung-Chul Baik; Min-Kyung Shin; Myunghwan Jung; Jin-Sik Park; Jong-Hoon Ha; Dong-Hae Lee; Min-Jeong Kim; Jeong-ih Shin; Hyung-Lyun Kang

doi:10.4167/jbv.2019.49.4.191

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Lee, Baik, Shin, Jung, Park, Ha, Lee, Kim, Shin, and Kang: Proteome Analysis of Alkylhydroxide Peroxidase-Deficient Isogenic Mutant of Helicobacter pylori 26695

Original Article

Journal of Bacteriology and Virology 2019; 49(4): 191-202.

Published online: 31 December 2019

DOI: https://doi.org/10.4167/jbv.2019.49.4.191

Proteome Analysis of Alkylhydroxide Peroxidase-Deficient Isogenic Mutant of Helicobacter pylori 26695

Woo-Kon Lee^1,², Seung-Chul Baik^1,², Min-Kyung Shin^1,², Myunghwan Jung^1,², Jin-Sik Park¹, Jong-Hoon Ha¹, Dong-Hae Lee¹, Min-Jeong Kim¹, Jeong-ih Shin¹, Hyung-Lyun Kang^1,²

¹Department of Microbiology, Gyeongsang National University School of Medicine, 816-15 Jinjudaero, Jinju, Gyeongnam, 52727 KOREA.

²Research Institute of Life Science, Gyeongsang National University 501 Jinjudaero, Jinju, Gyeongnam, 660-751 KOREA.

Corresponding: Hyung-Lyun Kang. Department of Microbiology, Gyeongsang National University School of Medicine, 816-15 Jinju-daero, Jinju, Gyeongnam, 52727 Korea. Phone: +82-55-772-8085, Fax: +82-55-772-8089, kangssi@gnu.kr

Received 5 December 2019 Revised 13 December 2019 Accepted 14 December 2019

The Korean Society for Microbiology and The Korean Society of Virology

(open-access):

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/).

Abstract

In order to investigate the antioxidant effect of alkylhydroxide peroxidase (ahpC) of Helicobacter pylori (H. pylori) 26695, an ahpC-deficient mutant (H. pylori 26695 ahpC::cat) was generated. ahpC-deficient mutant was grown slowly at lower pressure of oxygen (5% oxygen) compared to the H. pylori 26695. Whole cell proteins isolated form H. pylori 26695 and H. pylori 26695 ahpC::cat were analyzed by MALDI-TOF and tandem-MS. The expression of 15 proteins, including Ppa, HypB, GrpE, Elp, RecA, GroES, Mda66, RibE, NapA, GlnA, BioB, TrxB, Tsf, FumC and Icd, was more than doubled in H. pylori 26695 ahpC::cat. Production of 10 proteins such as UreG, FabE, Adk, Pnp, OorC, AtpA, AtpD, Nqq3, Pfr, and TagD decreased below 50% in H. pylori 26695 ahpC::cat compared to the H. pylori 26695. In microarray analysis, 9 genes including sul1, amiE, frxA, fecA, hyuA, and katA increased in transcription level in H. pylori 26695 ahpC::cat compared to H. pylori 26695. A total of 24 genes, including flaB, protein kinase C inhibitor, cag16, pabC, and sabA, reduced in transcription. 27 genes, including HP0889, showed common expression changes in ahpC, katA, and sodB-deficient mutations. As a result of this study, there were not many genes whose expression was commonly changed by the deletion of each of the three major antioxidant enzymes of H. pylori. These results showed the functions and regulation of the three antioxidant enzymes were different in H. pylori.

Keywords: Helicobacter pylori, antioxidant, ahpC

Figures and Tables

Figure 1

Strategy for construction of knock-out mutants (H. pylori 26695 ahpC::cat)

Figure 2

2D Separation of whole cell proteins of the wild-type (H. pylori 26695) (A), H. pylori 26695 ahpC::cat (B). The proteins were separated on an IPG strip of pH 5.0-8.0 and subsequently on a 12.5% SDS-PAGE and then detected by silver staining. The original gel size was 18×20×0.15. A) Wild type. B) H. pylori 26695 ahpC::cat. The protein spots corresponding to AhpC are marked with rectangle. kDa shows the molecular mass (indicated on left).

Figure 3

Growth curve of H. pylori 26695 and H. pylori 26695 ahpC::cat at thin layer culture condition.

Table 1

Strains and plasmids used in this study

Table 2

List of up- or down-regulated proteins in ahpC-deficient isogenic mutant compared to H. pylori 26695 by using tandem ms.

Table 3

List of up- or down-regulated proteins in ahpC-deficient isogenic mutant compared to H. pylori 26695 by using MALDI-TOF.

Table 4

List of genes of which expression are increased over two-fold in ahpC isogenic mutant when compared with wild type H. pylori 26695

Table 5

List of genes with increased or decreased expression common to ahpC, sodB, katA isogenic mutants when compared with H. pylori 26695

ACKNOWLEDGMENTS

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2016R1A2B1015791).

Notes

No potential conflict of interest relevant to this article was reported.

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