# Temporal and Spatial Profiling of Escherichia coli O157:H7 Surface Proteome: Insights into Intestinal Colonisation Dynamics In Vivo

## TL;DR

This study identifies surface proteins of E. coli O157:H7 during intestinal infection, revealing how they adapt over time and space to colonize the gut.

## Contribution

The first in vivo characterization of the surface proteome of E. coli O157:H7 during intestinal infection, revealing dynamic adaptations.

## Key findings

- 272 surface proteins were identified, with only 13 shared across all conditions, showing niche-specific adaptations.
- Proteomic shifts indicated a transition from stress adaptation to nutrient acquisition and persistence during infection.
- Cytoplasmic moonlighting proteins with surface activity were identified, suggesting novel roles in colonization.

## Abstract

Background: EHEC O157:H7 causes severe gastrointestinal illness by first colonizing the large intestine. It intimately attaches to the epithelial lining, orchestrating distinctive “attaching and effacing” lesions that disrupt the host’s cellular landscape. While much is known about the well-established virulence factors, there are much to learn about the surface proteins’ roles in a living host. Methods: This study presents the first in vivo characterisation of the surface proteome, i.e., proteosurfaceome, of Escherichia coli O157:H7 EDL933 during intestinal infection, revealing spatial and temporal adaptations critical for colonisation and survival. Using a murine ileal loop model, surface proteomic profiles were analysed at early (3 h) and late (10 h) infection stages across the ileum and colon. Results: In total, 272 proteins were identified, with only 13 shared across all conditions, reflecting substantial niche-specific adaptations. Gene ontology enrichment analyses highlighted dominant roles in metabolic, cellular, and binding functions, while subcellular localisation prediction uncovered cytoplasmic moonlighting proteins with surface activity. Comparative analyses revealed dynamic changes in protein abundance. Conclusions: These findings indicate a coordinated shift from stress adaptation and virulence to nutrient acquisition and persistence and provide a comprehensive view of EHEC O157:H7 surface proteome dynamics during infection, highlighting key adaptive proteins that may serve as targets for future therapeutic and vaccine strategies.