Proteomic Profiling of exosomes in cell culture supernatant of hBMEC infected by E. coli

ABSTRACT: Exosomes are small vesicles with a diameter of approximately 30-140nm. This double-layer phospholipid membrane encapsulates a variety of proteins, lipids and nucleic acids, which are produced and released by cells under specific conditions, participating in a large scale of regulatory processes. There is accumulating evidence indicating that exosomes also play an indispensable regulatory function in the central nervous system under physiological and pathological conditions, such as promoting communication between various cells in the blood-brain barrier. In our study, we attempted to obtain exosomes of human brain microvascular endothelial cells (hBMEC) infected by an extraintestinal pathogenic Escherichia. coli strain as well as control cells, and mass spectrometry analysis was performed to study the role of encapsulated proteome in central nervous system inflammation caused by bacterial infection. The particle size analysis results show that the size of the exosomes we isolated is mainly distributed between 30-200nm. Tetraspanin TSG101, CD9, and cytoplasmic protein Alix that we selected as positive markers for exosomes were successfully detected, while the negative marker, the endoplasmic reticulum-specific expression protein GRP94, was not found. In summary, the proteomic identification results showed that 1162 common proteins were identified in the two groups of exosomes from control and infected hBMEC, including 59 differentially expressed proteins. (Proteins with expression change more than 2 times or less than 0.5 times and P value less than 0.05 are regarded as differentially expressed proteins). The bioinformatics analysis indicated that differentially expressed proteins are mainly classified into structural molecular activity and constituent of ribosome. KEGG pathway enrichment analysis showed that many proteins are involved in ribosome-related pathways, pathogenic E. coli infections, and PI3K-Akt Signal pathways, etc. The present study laid an important foundation for the subsequent study of exosomal proteins in bacterial infections.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Brain Microvascular Endothelial Cell, Cell Culture

DISEASE(S): Bacterial Meningitis

SUBMITTER: Dong Huo  

LAB HEAD: Xiangru Wang

PROVIDER: PXD021064 | Pride | 2023-03-08

REPOSITORIES: Pride