SOLUBILIZATION OF ILEAL RECEPTOR FOR INTRINSIC FACTOR VITAMIN-B12 COMPLEX IN RAT

  • 1 January 1977
    • journal article
    • research article
    • Vol. 89  (1) , 172-180
Abstract
Solubilization of the ileal receptor for intrinsic factor-B12 complex (IF-B12) was attempted by extracting ileal mucosa of the rat with an alkaline buffer of pH 10 and mechanical grinding. The ileal extract, when incubated with homologous IF-57CoB12 and applied on a bio-Gel A-5m column, produced a macromolecular fraction containing IF-57CoB12, which was presumed to be a complex of receptor (Rec) and IF-B12. Ileal extract after centrifugation at 100,000 g for 1 h still yielded Rec-IF-B12. Formation of this complex in vitro was demonstrated by agar-gel electrophoresis in which a new peak appeared near the origin. This fraction was in the void volume of a Sephadex G-200 column. Ileal extracts obtained 1.5 and 3 h after oral administration of 57CoB12 contained Rec-IF-57CoB12. Treatment of Rec-IF-B12 with EDTA or trypsin released IF-B12, suggesting involvement of divalent cations in Rec-IF-B12 and a protein nature of Rec.

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