Proteomic analysis of the interaction of Bifidobacterium longum NCC2705 with the intestine cells Caco-2 and identification of plasminogen receptors.
To identify proteins with a potential role in the interaction of Bifidobacterium longum with intestinal epithelial cells, we profiled the protein response of B. longum NCC2705 following interaction with Caco-2 cells. Thirty-one protein spots, belonging to a total of 23 proteins, which exhibited a change in abundance of at least 3-fold were identified in B. longum NCC2705 following co-culture with Caco-2 cells, and were subsequently identified. Changes in expression were confirmed at the transcriptional level for a selection of these proteins. Enolase (Eno) and elongation factor Tu (EF-Tu) were amongst the proteins that showed the most prominent increase in abundance. Interaction of these proteins with plasminogen (Plg) was analyzed by Plg overlay assays, glutathione S-transferase (GST)-pull down, and western blot analysis. The results suggested that EF-Tu and Eno serve as surface receptors for B. longum NCC2705 binding to human plasminogen. Purified GST-EF-Tu and GST-Eno inhibited adhesion of B. longum NCC2705 to Caco-2 cells. Collectively, our data suggest that Eno and EF-Tu moonlight as adhesions, and are possibly involved in the protective role played by B. longum NCC2705 in defense against enteric pathogens. Biological significance The interaction of bifidobacteria with the human host plasminogen/plasmin system confirms the existence of a new component in the molecular cross-talk between bacteria and the host. Our study analyzed proteins EF-Tu and Eno with Plg binding activity, and they can inhibit adhesion of B. longum NCC2705 to Caco-2 cells, suggesting their role in the bacterial adherent to the enterocyte surface.
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Xiao
X
Wei
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Xiabei
X
Yan
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Xuannan
X
Chen
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Zhan
Z
Yang
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Huan
H
Li
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Dayang
D
Zou
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Xiang
X
He
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Simiao
S
Wang
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Qian
Q
Cui
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Wei
W
Liu
Institute of Microbiology and Biotechnology, University of Ulm, D-89069 Ulm, Germany.
Daria
D
Zhurina
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Xuesong
X
Wang
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Xiangna
X
Zhao
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China.
Liuyu
L
Huang
National Institutes for Food and Drug Control, 2 Tiantanxili, 100050 Beijing, China.
Ming
M
Zeng
Beijing Institute of Biotechnology, No. 27 TaiPing Lu Rd., 100850 Beijing, China. Electronic address: yeqn66@yahoo.com.
Qinong
Q
Ye
Institute of Microbiology and Biotechnology, University of Ulm, D-89069 Ulm, Germany. Electronic address: christian.riedel@uni-ulm.de.
Christian U
CU
Riedel
Institute of Disease Control and Prevention, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District 100071, Beijing, China. Electronic address: yuanjing6216@163.com.
Jing
J
Yuan
D001422:Bacterial Adhesion / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D001644:Bifidobacterium / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D018938:Caco-2 Cells; D018920:Coculture Techniques; D006801:Humans; D007413:Intestinal Mucosa / Q000276:immunology / Q000378:metabolism* / Q000382:microbiology; D010444:Peptide Elongation Factor Tu / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D010751:Phosphopyruvate Hydratase / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D010958:Plasminogen / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D040901:Proteomics
D001426:Bacterial Proteins; D010958:Plasminogen; D010444:Peptide Elongation Factor Tu; D010751:Phosphopyruvate Hydratase
Adhesion; Bifidobacterium; EF-Tu; Enolase; Plasminogen