Bacterial response to eukaryotic cells. Analysis of differentially expressed proteins using nano liquid chromatography-electrospray ionization tandem mass spectrometry.
Host-bacteria interactions have mostly been investigated with regard to the host response or to activities of pathogenic bacteria. In contrast, we aim to identify reactions of non-pathogenic bacteria that result from their contact with host cells of the gastrointestinal tract. In a proteomic approach, the response of non-pathogenic human Escherichia coli bacteria on gut epithelial cells (rat IEC-6) was investigated in an in vitro co-culture model. For this purpose, a sensitive analytical procedure was developed based on the identification of two-dimensional polyacrylamide gel electrophoresis separated proteins by online nanoLC-electrospray ionization MS/MS using a quadrupole time-of-flight tandem mass spectrometer for accurate mass determination. We demonstrate here the efficiency of this technique by the identification of a total of 43 differentially expressed proteins, out of which 25 were up-regulated and 18 were down-regulated. They represent a wide range of molecular weight and different metabolic and physiological functions.
Alpert
C
C
German Institute of Human Nutrition Potsdam-Rehbruecke, Department of Gastrointestinal Microbiology, A.-Scheunert-Allee 114-116, D-14558 Nuthetal, Germany. alpert@mail.dife.de
Engst
W
W
Guehler
A
A
Oelschlaeger
T
T
Blaut
M
M
D000818:Animals; D002460:Cell Line; D002853:Chromatography, Liquid / Q000379:methods*; D018920:Coculture Techniques; D015536:Down-Regulation; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis*; D007413:Intestinal Mucosa / Q000166:cytology / Q000382:microbiology*; D036103:Nanotechnology / Q000379:methods; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D015854:Up-Regulation
D029968:Escherichia coli Proteins