TMT-Based Quantitative Proteomic Analysis of Intestinal Organoids Infected by Listeria monocytogenes with Different Virulence

Abstract

Listeria monocytogenes (Lm) is an opportunistic food-borne pathogen that cause listeriosis. L. monocytogenes belonged to different serovars presents with different virulence in the host and caused different host reactions. To investigate the remodeling of host proteome by differently toxic strains, the cellular protein responses of intestinal organoids were analyzed using TMT labeling and high performance liquid chromatography-mass spectrometry. Quantitative proteomic analysis revealed 6564 differentially expressed proteins, of which 5591 proteins were quantified. The fold-change cutoff was set at 1.3 (Lm vs control), the virulent strain caused 102 up-regulated proteins and 52 down-regulated proteins, while the low virulent strain caused 188 up-regulated proteins and 25 down-regulated proteins. These identified proteins were involved in the regulation of essential processes such as biological metabolism, energy metabolism, and immune system process. Some selected proteins were screened by Real-time PCR and Western blotting. These results revealed that differently toxic L. monocytogenes induced similar biological functions and immune responses while had different regulation on differential proteins in the pathway.