A recombinant catalytic domain of matriptase induces detachment and apoptosis of small-intestinal epithelial IEC-6 cells cultured on laminin-coated surface.
Matriptase is a type-II transmembrane serine protease that is expressed strongly in the epithelial elements of various organs. In the small intestine, it is expressed prominently at the villus tip where aged epithelial cells undergo shedding and/or apoptosis. This observation, together with the ability of matriptase to cleave laminin (a basement membrane component critical for epithelial cell attachment), prompted us to hypothesize that it plays an important part in the removal of aged epithelial cells in the small intestine. We tested this hypothesis by determining whether a recombinant catalytic domain of rat matriptase (His(6)t-S-CD) causes detachment and/or apoptosis of small-intestinal epithelial IEC-6 cells. His(6)t-S-CD caused detachment of cells attached to laminin-coated plates but did not detach cells attached to fibronectin- or type-IV collagen-coated plates. Pre-treatment of laminin-coated plates with His(6)t-S-CD decreased the attachment of cells, suggesting that the recombinant matriptase caused detachment through a mechanism involving a direct effect on laminin. His(6)t-S-CD was also found to induce apoptosis in the cells cultured on laminin-coated plates, as assessed by annexin-V staining, DNA fragmentation and caspase-3 activity assays. These findings support our hypothesis regarding the role of matriptase in the small intestine.
Mochida
Seiya
S
Laboratory of Nutrition Chemistry, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto, Japan.
Tsuzuki
Satoshi
S
Inouye
Kuniyo
K
Fushiki
Tohru
T
D000818:Animals; D017302:Annexins / Q000032:analysis; D017209:Apoptosis / Q000502:physiology; D053148:Caspase 3 / Q000032:analysis; D020134:Catalytic Domain / Q000502:physiology; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003001:Cloning, Molecular; D053938:DNA Fragmentation / Q000187:drug effects; D006639:Histidine / Q000235:genetics / Q000378:metabolism; D007413:Intestinal Mucosa; D007421:Intestine, Small / Q000502:physiology; D007797:Laminin / Q000737:chemistry* / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism; D010843:Pichia; D011480:Protease Inhibitors / Q000494:pharmacology; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism
D017302:Annexins; C471213:His-His-His-His-His-His; D007797:Laminin; D008565:Membrane Proteins; D009842:Oligopeptides; D011480:Protease Inhibitors; D020033:Protein Isoforms; D011994:Recombinant Proteins; D006639:Histidine; D012697:Serine Endopeptidases; C467801:matriptase 2; D053148:Caspase 3