Identification of Functional Interactome of Gastric Cancer Cells with Helicobacter pylori Outer Membrane Protein HpaA
Ruyue Fan1, Xiurui Han1, Di Xiao1
1State Key Laboratory of Infectious Disease Prevention and Control, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
Abstract:
HpaA as an outer membrane protein of Helicobacter pylori (H. pylori) plays a significant role in the adhesion to the human stomach, but the functional relation between HpaA and gastric epithelial cells is still not clear. To screen the interaction between HpaA and cellular proteins in gastric epithelial cells, the HpaA protein from H. pylori 26695 fused with a tag (6× His) was expressed and purified successfully, the secondary structure was estimated by the Circular Dichroism (CD) spectrum, and the purified recombinant protein was used to perform the pull-down assays with gastric cancer cell lines (AGS and SGC-7901) lysates, respectively. The pull-down proteins were identified by high-performance liquid chromatography tandem mass spectrometry system (HPLC-MS/MS). A total of 9 and 13 proteins related were analyzed from AGS and SGC-7901 cell lysates, respectively. ANXA2 was considered as putative HpaA functional partner discovered from lysates of both cell lines with high score and coverage. It is hypothesized that HpaA may be involved in the biological process of regulation of transcription and nucleic acid metabolism during the adhesion of H. pylori to human gastric epithelial cells, and HpaA-binding proteins also be used as targets for the development of antiadhesion drugs against H. pylori.
Insights
Helicobacter pylori outer membrane protein HpaA interacts with gastric epithelial cells. Researchers identified ANXA2 as a key binding partner, suggesting new targets for anti-adhesion drugs.
Area of Science:
- Microbiology
- Molecular Biology
- Cell Biology
Background:
- Helicobacter pylori (H. pylori) outer membrane protein HpaA is crucial for bacterial adhesion to the human stomach.
- The precise functional relationship between HpaA and gastric epithelial cells remains unclear.
Purpose of the Study:
- To investigate the interactions between HpaA and cellular proteins in gastric epithelial cells.
- To identify potential HpaA binding partners involved in H. pylori adhesion.
Main Methods:
- Recombinant HpaA protein from H. pylori 26695 was expressed, purified, and characterized using Circular Dichroism (CD) spectroscopy.
- Pull-down assays were performed using purified HpaA and lysates from gastric cancer cell lines (AGS and SGC-7901).
- Proteins interacting with HpaA were identified using high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS).
Main Results:
- A total of 9 and 13 proteins were identified from AGS and SGC-7901 cell lysates, respectively.
- Annexin A2 (ANXA2) was consistently identified as a high-scoring putative HpaA functional partner in both cell lines.
- HpaA may be involved in regulating transcription and nucleic acid metabolism during H. pylori adhesion.
Conclusions:
- ANXA2 is a potential functional partner of HpaA in gastric epithelial cells.
- HpaA-binding proteins, such as ANXA2, represent potential therapeutic targets for developing anti-adhesion drugs against H. pylori infections.
