Identification of inflammation-related proteins in a murine colitis model by 2D fluorescence difference gel electrophoresis and mass spectrometry.
OBJECTIVE
The aim of this study was to identify new intestinal proteins potentially associated with acute inflammation using proteomic profiling of an in vivo mice model of ulcerative colitis.


METHODS
2D fluorescence difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization time-of-flight spectrometer (MALDI-TOF) peptide mass fingerprinting were used to determine differentially expressed proteins between normal and inflamed intestinal mucosa. Acute colitis was induced by 8.0% dextran sodium sulfate (DSS) given p.o. for 7 days.


RESULTS
Among a total of seven protein spots showing differential expression, we identified five different proteins, of which two were upregulated and three downregulated in colitis in comparison to normal mucosa, using the MASCOT search engine. 3-Hydroxy-3-methylglutaryl-coenzyme A synthase 2 and serpin b1a were upregulated proteins, and protein disulfide-isomerase A3, peroxiredoxin-6 and vimentin were identified as downregulated proteins.


CONCLUSIONS
These identified proteins may be responsible for the development of the intestinal inflammation. 2D-DIGE and MALDI-TOF mass spectrometry are useful in the search for the differentially expressed proteins.
Naito
Yuji
Y
Molecular Gastroenterology and Hepatology, Graduate School of Medical Science, Kyoto, Japan. ynaito@koto.kpu-m.ac.jp
Takagi
Tomohisa
T
Okada
Hitomi
H
Omatsu
Tatsushi
T
Mizushima
Katsura
K
Handa
Osamu
O
Kokura
Satoshi
S
Ichikawa
Hiroshi
H
Fujiwake
Hideshi
H
Yoshikawa
Toshikazu
T
D000818:Animals; D003093:Colitis, Ulcerative / Q000139:chemically induced / Q000378:metabolism*; D003106:Colon / Q000378:metabolism*; D016264:Dextran Sulfate; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D005453:Fluorescence; D006904:Hydroxymethylglutaryl-CoA Synthase / Q000378:metabolism; D018836:Inflammation Mediators / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010449:Peptide Mapping; D054465:Peroxiredoxin VI / Q000378:metabolism; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D011506:Proteins / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D015843:Serpins / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014746:Vimentin / Q000378:metabolism
D018836:Inflammation Mediators; D011506:Proteins; C522950:Serpinb1a protein, mouse; D015843:Serpins; D014746:Vimentin; D016264:Dextran Sulfate; D054465:Peroxiredoxin VI; C515983:Prdx6 protein, mouse; D006904:Hydroxymethylglutaryl-CoA Synthase; C513195:Pdia3 protein, mouse; D019704:Protein Disulfide-Isomerases