2-DE and MS analysis of interactions between Lactobacillus fermentum I5007 and intestinal epithelial cells.
Lactobacillus is a probiotic commonly used for supplementation to human and animal diets. In this study, we used 2-DE and MS to analyze changes in the proteomes of Lactobacillus and intestinal epithelial cells in two model systems. The in vivo and in vitro models were involved the inoculation of Lactobacillus fermentum I5007 into the rabbit jejunum for 4 h and the culture of the bacterium with Caco-2 cells for 1 h, respectively. Our results indicate that, after exposure to the intestinal environment, the bacterium exhibited decreases in key enzymes involved in energy metabolism (e.g., lactate dehydrogenase, dihydrolipoamide dehydrogenase, and nicotinate phosphoribosyltransferase) and amino acid metabolism (e.g., arginyl-tRNA synthetase and aspartate-semialdehyde dehydrogenase), but increases in glycoside hydrolase (an enzyme for mucin degradation) and fructose-6-phosphate phosphoketolase (an enzyme of the pentose phosphate pathway). In response to an interaction with L. fermentum I5007, Caco-2 cells showed changes in proteins that were beneficial for gut integrity, including voltage-dependent anion channel 1, glutathione transferase, and heat shock protein gp96. On the basis of their functions, we suggest that these proteins serve as useful biomarkers for metabolic changes in Lactobacillus and intestinal epithelial cells in response to their interactions.
Yang
Fang
F
National Key Laboratory of Animal Nutrition, China Agricultural University, Beijing, PR China.
Wang
Junjun
J
Li
Xiaojie
X
Ying
Tianyi
T
Qiao
Shiyan
S
Li
Defa
D
Wu
Guoyao
G
D000818:Animals; D000951:Antigens, Neoplasm / Q000032:analysis / Q000378:metabolism; D001422:Bacterial Adhesion; D015415:Biomarkers / Q000032:analysis; D018938:Caco-2 Cells / Q000378:metabolism; D002450:Cell Communication; D002457:Cell Extracts / Q000032:analysis; D018920:Coculture Techniques / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004734:Energy Metabolism; D005982:Glutathione Transferase / Q000032:analysis / Q000378:metabolism; D006801:Humans; D007413:Intestinal Mucosa / Q000737:chemistry / Q000378:metabolism / Q000382:microbiology*; D007583:Jejunum / Q000166:cytology / Q000382:microbiology; D052197:Limosilactobacillus fermentum / Q000737:chemistry / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000032:analysis; D019936:Probiotics / Q000494:pharmacology; D020543:Proteome / Q000032:analysis; D011817:Rabbits; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D050995:Voltage-Dependent Anion Channel 1 / Q000032:analysis / Q000378:metabolism
D000951:Antigens, Neoplasm; D015415:Biomarkers; D002457:Cell Extracts; D009363:Neoplasm Proteins; D020543:Proteome; C052646:sarcoma glycoprotein gp96 rejection antigens; D050995:Voltage-Dependent Anion Channel 1; D005982:Glutathione Transferase