CEL-I, an N-acetylgalactosamine (GalNAc)-specific C-type lectin, induces nitric oxide production in RAW264.7 mouse macrophage cell line.
We found that CEL-I, a GalNAc-specific C-type lectin isolated from the marine invertebrate Holothuroidea (Cucumaria echinata), induces inducible nitric oxide synthase (iNOS) expression and NO production in RAW264.7 cells. The NO production was inhibited by an iNOS inhibitor, L-NAME, but was not by a lipopolysaccharide (LPS) inhibitor, polymyxin B. In the presence of 0.1-M GalNAc, increased NO production by CEL-I-treated RAW264.7 cells was observed rather than the inhibition. Bovine serum albumin (BSA) significantly inhibited the CEL-I-induced NO production as well as the binding of FITC-labelled CEL-I on RAW264.7 cells. Three MAP kinase inhibitors (specific to extra-cellular regulated kinase, c-jun NH(2)-terminal kinase and p38 MAP kinase) inhibited CEL-I-induced NO production with different extents. Heat-treatment of CEL-I resulted in a decreased activity of CEL-I depending on the temperature. These results suggest that CEL-I induces NO production in RAW264.7 cells through the protein-cell interaction rather than the binding to the specific carbohydrate chains on the cell surface.
Nagasaki University, Bunkyo-machi, Japan.
Tomohiro
T
Yamanishi
Tomomitsu
T
Hatakeyama
Kenichi
K
Yamaguchi
Tatsuya
T
Oda
D000116:Acetylgalactosamine / Q000737:chemistry*; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D048016:Cucumaria / Q000737:chemistry*; D005786:Gene Expression Regulation; D037181:Lectins, C-Type / Q000737:chemistry* / Q000378:metabolism*; D008264:Macrophages / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D010835:Phytohemagglutinins / Q000737:chemistry / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014909:Wheat Germ Agglutinins / Q000737:chemistry / Q000494:pharmacology
D037181:Lectins, C-Type; D010835:Phytohemagglutinins; D014909:Wheat Germ Agglutinins; C100156:leukoagglutinins, plants; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; D000116:Acetylgalactosamine