Cubilin expression and posttranslational modification in the canine gastrointestinal tract.
Cubilin is an endocytic receptor of the apical brush border membrane that is essential for intrinsic factor-mediated cobalamin absorption in small intestine. However, cubilin is more highly expressed in kidney and yolk sac, and recent molecular characterization of the receptor has focused on these tissues. The aim of this investigation was to examine tissue-specific cubilin expression and posttranslational modifications with an emphasis on the gastrointestinal tract. Intrinsic factor-cobalamin binding activity, cubilin immunoreactivity, and cubilin mRNA levels were determined in multiple segments of canine gastrointestinal mucosa and other tissues. These aspects of cubilin expression varied in parallel, suggesting that the major determinant of regional cubilin expression in the gastrointestinal tract is modulation of cubilin mRNA. Cell fractionation indicated that ileal cubilin is not strongly membrane associated. An approximately 185-kDa brush border specific and two >400-kDa precursor forms of cubilin were identified. Asparagine-linked oligosaccharide modifications characterized by differential glycosidase digestion of affinity-purified cubilin from ileal mucosa and renal cortex differed, but ileal and renal intracellular cubilin comigrated on SDS-PAGE at approximately 400 kDa after oligosaccharide removal, thus reconciling previous conflicting size estimates of the cubilin polypeptide.
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Department of Microbiology and Small Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, Michigan 48824, USA.
Fyfe
J C
JC
D000818:Animals; D003037:Cobalt Radioisotopes / Q000493:pharmacokinetics; D003106:Colon / Q000378:metabolism; D004285:Dogs; D004947:Esophagus / Q000378:metabolism; D005753:Gastric Mucosa / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000378:metabolism*; D007421:Intestine, Small / Q000378:metabolism; D007668:Kidney / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008871:Microvilli / Q000378:metabolism; D009928:Organ Specificity; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000235:genetics; D011869:Radioligand Assay; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic; D014805:Vitamin B 12 / Q000378:metabolism
D003037:Cobalt Radioisotopes; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; C029486:intrinsic factor-cobalamin receptor; D014805:Vitamin B 12