Characterization of a putative receptor for intestinal trefoil factor in rat small intestine: Identification by in situ binding and ligand blotting
Characterization of a putative receptor for intestinal trefoil factor in rat small intestine: Identification by in situ binding and ligand blotting
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DOI：
10.1006/bbrc.1997.7144
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发表时间：
1997-08-28
影响因子：
3.1
通讯作者：
GonzalezCrussi, F
中科院分区：
文献类型：
--
作者：
Tan, XD;Hsueh, W;GonzalezCrussi, F
肠三叶因子（ITF）是一种由肠道杯状细胞分泌的小肽，可维持黏膜完整性并促进上皮伤口愈合。尽管ITF已被克隆，但其与肠上皮相互作用的详细机制仍不清楚。在本研究中，我们利用pXa（一种原核表达载体）在大肠杆菌中表达成熟大鼠ITF（rITF），以产生一种生物素化的rITF融合蛋白（bTag - ITF）。通过使用bTag - ITF探针，我们在小肠隐窝和胃腺区域的黏液细胞中鉴定出ITF结合细胞。利用配体印迹技术，我们进一步从肠黏膜细胞膜组分中鉴定出一种50kDa的糖蛋白，它可与bTag - ITF结合。我们的数据表明，这种50kDa的膜糖蛋白是胃肠道中ITF的一种假定受体。（C）1997年学术出版社。
Intestinal trefoil factor (ITF), a small peptide secreted by intestinal goblet cells, maintains mucosal integrity and promotes epithelial wound healing. Although ITF has been cloned, the detailed mechanism by which ITF interacts with intestinal epithelium remains elusive. In the present study, we expressed mature rat ITF (rITF) with pXa (a prokaryotic expression vector) in Escherichia coli to generate a biotinylated rITF fusion protein (bTag-ITF). By using bTag-ITF probe, we identified ITF binding cells in the crypts of the small intestine and mucous cells of the region of gastric glands. Using a ligand blotting technique, we further characterized a 50 kDa glycosylated protein from the membrane fraction of the small intestine, which bound to bTag-ITF. Our data suggest that this 50 kDa membrane glycoprotein is a putative receptor for ITF in the gastrointestinal tract. (C) 1997 Academic Press.