Related Experiment Video
Updated: Jun 7, 2025
Experimental Protocol for Detecting Mitochondrial Function in Hepatocytes Exposed to Organochlorine Pesticides
Published on: September 16, 2020
The Hcp2b of APEC induces mitochondrial damage in chicken DF-1 cells
Liting Lu1,2,3, Zhao Qi1,2,3, Haiyang Wang1,2,3
1Joint Research Center for Food Nutrition and Health of IHM, Anhui Agricultural University, Hefei, People's Republic of China.
Abstract:
The haemolysin co-regulatory protein (Hcp) plays a significant role in the pathogenicity of avian pathogenic Escherichia coli (APEC) as an effector protein of the type VI secretion system (T6SS) to the host. Meanwhile, mitochondria in the host are the target of effector proteins of various secretion systems. Here, we explored the effects of APEC effector Hcp2b on the mitochondria of DF-1 cells and found that Hcp2b results in damage in mitochondria. Next, 68 target proteins in DF-1 cell lysates were identified that interacted with Hcp2b by streptavidin-biotin pull-down assay combined with LC-MS/MS, among which ADP/ATP transporter carrier (SLC25A4) is a mitochondria-associated protein; protein docking analysis showed that Hcp2b binds well to SLC25A4. Therefore, we hypothesize that the Hcp2b contributes to mitochondrial damage in DF-1 cells through interaction with the SLC25A4.
Insights
Avian pathogenic Escherichia coli (APEC) effector Hcp2b damages host mitochondria. This study identifies ADP/ATP transporter SLC25A4 as a binding partner, suggesting Hcp2b causes mitochondrial injury via SLC25A4 interaction.
Area of Science:
- Microbiology
- Cell Biology
- Pathogenesis
Background:
- Avian pathogenic Escherichia coli (APEC) utilizes effector proteins like haemolysin co-regulatory protein (Hcp) to cause disease.
- Mitochondria are critical cellular organelles frequently targeted by bacterial virulence factors.
Purpose of the Study:
- To investigate the impact of APEC Hcp2b on host cell mitochondria.
- To identify host proteins interacting with Hcp2b and elucidate the mechanism of mitochondrial damage.
Main Methods:
- Exposure of DF-1 cells to APEC Hcp2b.
- Streptavidin-biotin pull-down assay coupled with LC-MS/MS to identify Hcp2b interacting proteins.
- Protein-protein docking analysis.
Main Results:
- APEC Hcp2b induced significant mitochondrial damage in DF-1 cells.
- Sixty-eight potential Hcp2b interacting proteins were identified, including ADP/ATP transporter carrier (SLC25A4).
- Docking analysis confirmed a strong binding affinity between Hcp2b and SLC25A4.
Conclusions:
- APEC Hcp2b directly damages host cell mitochondria.
- The interaction between Hcp2b and SLC25A4 is hypothesized to mediate APEC-induced mitochondrial dysfunction.
