J Bacteriol Virol.  2010 Jun;40(2):67-75. 10.4167/jbv.2010.40.2.67.

Analysis of Low Molecular Weight Proteome from H. pylori Cell Extract Using the High Performance Liquid Chromatography

• Park JW
• Lee KJ
• Kim KM
• Joo JS
• Kwon YC
• Youn HS
• Song JY
• Kang HL
• Lee KH
• Baik SC
• Lee WK
• Cho MJ
• Rhee KH

• KMID: 1456269
• DOI: http://doi.org/10.4167/jbv.2010.40.2.67

Abstract

Low molecular proteins (LMPs) which are smaller than 20 kDa are difficult to visible on a standard two-dimensional SDS-polyacrylamide gel electrophoresis (2-D SDS-PAGE) map. LMPs must be enriched appropriately to be analyzed. We isolated LMPs of Helicobacter pylori 26695 from 1-D polyacrylamide gel and digested by pepsin. Pepsin-digested LMPs were separated by HPLC and each fraction was analyzed by hybrid tandem mass spectrometer. Seventy nine peptides, representing 27 genes, including copper ion binding protein (CopP, 7 kDa), thioredoxin (TrxA, 11.9 kDa) and ribosomal protein L23 (Rpl23, 10.5 kDa) were identified. Some proteins larger than 40 kDa including Omp2, Omp21, Omp27, Omp30, Omp32, catalase and HP1083 were also identified. This work may give researchers a useful way to analyse the expressed LMPs which could not be identified on the conventional 2-D SDS-PAGE.

Keyword

Helicobacter pylori; Hybrid tandem mass spectrometer; Low molecular protein

MeSH Terms

Acrylic Resins
Carrier Proteins
Catalase
Chimera
Chromatography, High Pressure Liquid
Chromatography, Liquid
Copper
Electrophoresis
Electrophoresis, Polyacrylamide Gel
Helicobacter pylori
Molecular Weight
Pepsin A
Peptides
Proteins
Proteome
Ribosomal Proteins
Thioredoxins
Acrylic Resins
Carrier Proteins
Catalase
Copper
Pepsin A
Peptides
Proteins
Proteome
Ribosomal Proteins
Thioredoxins

Figure

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  Figure 1. Extraction of the H. pylori LMW proteins. In lanes: M, molecular weight marker; 1, whole cell proteins of H. pylori 26695; 2, extracted low molecular proteins.

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  Figure 2. Profile of the H. pylori low molecular proteins separated by reverse phased HPLC.

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  Figure 3. 2-DE map of the low molecular proteins of H. pylori that were extracted from 1-DE gel.

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