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Identification of Mycoplasma pneumoniae P1-interacting proteins by GST pull-down and analysis of its transcriptomic
Chao Yan1, Xuanfeng Liu1, An Su1
1Department of Bacteriology, Capital Center for Children's Health, Capital Medical University, Capital Institute of Pediatrics, Beijing, 100020, China.
Abstract:
Mycoplasma pneumoniae (M. pneumoniae) is a major pathogen causing community-acquired pneumonia in children. Its pathogenic process relies on the adherence to and colonization of host respiratory epithelial cells. P1 protein is the primary adhesin of M. pneumoniae, directly mediating its binding to host cells. To explore the interaction mechanism between P1 recombinant protein and host cells, we conducted protein expression and purification, glutathione S-transferase (GST) pull-down assay, and transcriptome sequencing. The rP1-GST fusion protein was expressed under confirmed induction conditions (16°C, 0.1 mM IPTG). GST pull-down assay identified 22 differentially expressed membrane proteins in the rP1-GST group, among which annexin A2 (ANXA2) and C-C chemokine receptor type 5 (CCR5) were significantly altered and interact with P1 adhesin. Both ANXA2 and CCR5 possessed multiple functions including protein binding, receptor activity and signal sensor activity. Transcriptome analysis indicated that differentially expressed genes from rP1-A549 cell interaction were significantly enriched in multiple Gene Ontology (GO) terms and KEGG pathways. These results suggest that ANXA2 and CCR5 may serve as potential binding partners of P1 adhesin. P1 adhesin may regulate host genes involved in mitochondria and energy metabolism. These findings provide clues for understanding the adhesion and pathogenesis of M. pneumoniae.
Insights
Mycoplasma pneumoniae P1 adhesin binds to host cells via annexin A2 and CCR5. This interaction influences host genes related to mitochondria and energy metabolism, offering insights into M. pneumoniae pathogenesis.
Area of Science:
- Microbiology
- Molecular Biology
- Cell Biology
Background:
- Mycoplasma pneumoniae causes childhood pneumonia by adhering to respiratory cells.
- The P1 protein is crucial for M. pneumoniae attachment to host cells.
Purpose of the Study:
- To investigate the interaction mechanism between the M. pneumoniae P1 recombinant protein and host cells.
- Identify host cell binding partners of the P1 adhesin.
Main Methods:
- Recombinant P1-GST fusion protein expression and purification.
- Glutathione S-transferase (GST) pull-down assay to identify interacting proteins.
- Transcriptome sequencing to analyze host gene expression changes.
Main Results:
- GST pull-down identified 22 differentially expressed membrane proteins, including annexin A2 (ANXA2) and C-C chemokine receptor type 5 (CCR5) as P1 interactors.
- ANXA2 and CCR5 exhibit protein binding, receptor activity, and signal sensing functions.
- Transcriptome analysis revealed enrichment of Gene Ontology terms and KEGG pathways related to mitochondria and energy metabolism.
Conclusions:
- ANXA2 and CCR5 are potential binding partners for the M. pneumoniae P1 adhesin.
- P1 adhesin may modulate host genes involved in mitochondrial function and energy metabolism.
- Findings provide insights into M. pneumoniae adhesion and pathogenesis mechanisms.
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