J Bacteriol Virol.  2019 Dec;49(4):191-202. 10.4167/jbv.2019.49.4.191.

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

• Lee WK
• Baik SC
• Shin MK
• Jung M
• Park JS
• Ha JH
• Lee DH
• Kim MJ
• Shin JI
• Kang HL

• KMID: 2468025
• DOI: http://doi.org/10.4167/jbv.2019.49.4.191

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.

Keyword

Helicobacter pylori; antioxidant; ahpC

MeSH Terms

Antioxidants
Helicobacter pylori*
Helicobacter*
Microarray Analysis
Oxygen
Peroxidase
Protein Kinase C
Proteome*
Ribes
Antioxidants
Oxygen
Peroxidase
Protein Kinase C
Proteome

Figure

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  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.

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