Identification of a macromolecular factor in the ileum which binds intrinsic factor and immunologic identification of intrinsic factor in ileal extracts

AUTOR(ES)

Rothenberg, Sheldon P.

RESUMO

The precipitate which resulted when 57CoB12 bound to normal human gastric juice was subjected to a 15% concentration of Na2SO4 contained virtually no radioactivity. However, after in vivo incubation of the gastric juice-57CoB12 mixture in the distal ileum of the guinea pig, the dialyzed extract of the washed mucosa contained a fraction of 57CoB12 which was precipitated at 15% Na2SO4. In addition, in vitro incubation of gastric juice-57CoB12 with an extract of the ileal mucosa or brush border membranes also resulted in the formation of a 15% Na2SO4-insoluble fraction which contained 57CoB12. The formation of this 57CoB12-containing insoluble fraction did not occur or was diminished by (a) addition of an excess of B12-free normal human gastric juice. (b) reducing the incubation pH to 2, (c) incubating the mixture at 4°C, (d) pretreating the ileal extract at 56°C for 30 min, (e) incubating the reaction in sodium EDTA but not calcium EDTA, (f) incubating gastric juice-57CoB12 with an extract of jejunal mucosa. Sephadex gel filtration was used to demonstrate that the factor in the ileal extract which reacted with the gastric juice-57CoB12 filtered through G-100 and G-200 columns in the excluded volume.

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