Purification by cobalamin-Sepharose affinity chromatography and intrinsic factor-binding activity of an extramembrane proteolytic product from pig ileal mucosa
| Autor: | Jean-Louis Guéant, Amal Safi, Monique Saunier, Jean-Claude Michalski, Isabelle Gastin, Alibada Yerima |
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| Rok vydání: | 1996 |
| Předmět: |
Intrinsic Factor
Swine Biochemistry Cobalamin Chromatography Affinity Sepharose chemistry.chemical_compound Affinity chromatography Intestinal mucosa Ileum hemic and lymphatic diseases polycyclic compounds Animals heterocyclic compounds Intestinal Mucosa Molecular Biology Polyacrylamide gel electrophoresis Intrinsic factor integumentary system Molecular mass Hydrolysis Proteins nutritional and metabolic diseases Cell Biology Molecular biology Kinetics Vitamin B 12 Isoelectric point chemistry Chromatography Gel Autoradiography Electrophoresis Polyacrylamide Gel Protein Binding Research Article |
| Zdroj: | Biochemical Journal. 313:675-681 |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bj3130675 |
| Popis: | We have purified a cobalamin-binding protein obtained by papain digestion of pig intestine by cobalamin-AH-Sepharose affinity chromatography, with a purification factor of 17300, a yield of 63% and a cobalamin-binding activity of 11260 pmol/mg of protein. The protein contained 3.8% carbohydrate and was O- and N-glycosylated. Its molecular mass was 69 kDa on SDS/PAGE and its isoelectric point was 5.1. It had a binding activity for both [57Co]cobalamin and [57Co]cobalamin-intrinsic factor in native PAGE autoradiography and it inhibited the binding of intrinsic factor to the intact intestinal receptor with an IC50 of 49.31 nmol/l in a radioisotope assay. In conclusion, the purified protein shared a binding activity for both cobalamin and intrinsic factor-cobalamin complexes and could correspond to the extracellular domain of the ileal intrinsic factor receptor. |
| Databáze: | OpenAIRE |
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