Identification of a surface protein from Lactobacillus reuteri JCM1081 that adheres to porcine gastric mucin and human enterocyte-like HT-29 cells.
Adhesion of lactobacilli to the host gastrointestinal (GI) tract is considered an important factor in health-promoting effects. However, studies addressing the molecular mechanisms of the adhesion of lactobacilli to the host GI tract have not yet been performed. The aim of this work was to identify Lactobacillus reuteri surface molecules mediating adhesion to intestinal epithelial cells and mucins. Nine strains of lactobacilli were tested for their ability to adhere to human enterocyte-like HT-29 cells. The cell surface proteins involved in the adhesion of Lactobacillus to HT-29 cells and gastric mucin were extracted. The active fractions were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting with horseradish peroxidase-labeled mucin and NHS-Biotin-labeled HT-29 cells. Furthermore, tandem mass spectrometry analysis was performed to identify the surface protein that participates in adhesion. It was shown that the ability of lactobacilli to adhere to HT-29 cells in vitro varied considerably among different strains. The most adhesive strain was the chicken intestinal tract isolate Lactobacillus reuteri JCM1081 (495.07 +/- 80.03 bacterial cells/100 HT-29 cells). The adhesion of L. reuteri JCM1081 to HT-29 cells appeared to be mediated by a cell surface protein, with an approximate molecular mass of 29 kDa. The peptides generated from the 29-kDa protein significantly matched the Lr0793 protein sequence of L. reuteri strain ATCC55730 (~71.1% identity) and displayed significant sequence similarity to the putative ATP-binding cassette transporter protein CnBP.
Wang
Bin
B
Research Institute of General Surgery, Jin Ling Hospital, Nanjing, Jiangsu, China.
Wei
Hong
H
Yuan
Jing
J
Li
Qiurong
Q
Li
Yousheng
Y
Li
Ning
N
Li
Jieshou
J
D000818:Animals; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004847:Epithelial Cells / Q000382:microbiology; D005752:Gastric Mucins / Q000378:metabolism*; D019073:HT29 Cells; D006801:Humans; D007413:Intestinal Mucosa / Q000382:microbiology*; D052200:Limosilactobacillus reuteri / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008970:Molecular Weight; D013552:Swine
D001426:Bacterial Proteins; D005752:Gastric Mucins; D008565:Membrane Proteins