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.
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January 1996
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Research Article|
January 15 1996
Purification by cobalamin-Sepharose affinity chromatography and intrinsic factor-binding activity of an extramembrane proteolytic product from pig ileal mucosa Available to Purchase
Alibada YERIMA;
Alibada YERIMA
*Laboratoire de Biologie Cellulaire et Moléculaire en Nutrition et INSERM Unité 308 Faculté de Médecine, Université de Nancy, Nancy, France
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Amal SAFI;
Amal SAFI
*Laboratoire de Biologie Cellulaire et Moléculaire en Nutrition et INSERM Unité 308 Faculté de Médecine, Université de Nancy, Nancy, France
†Laboratoire de Biochimie Faculté de Sciences, Université de Meknès, Morocco
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Isabelle GASTIN;
Isabelle GASTIN
*Laboratoire de Biologie Cellulaire et Moléculaire en Nutrition et INSERM Unité 308 Faculté de Médecine, Université de Nancy, Nancy, France
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Jean-Claude MICHALSKI;
Jean-Claude MICHALSKI
‡Laboratoire de Chimie Biologique, Université de Lille I, Villeneuve d'Ascq, France
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Monique SAUNIER;
Monique SAUNIER
*Laboratoire de Biologie Cellulaire et Moléculaire en Nutrition et INSERM Unité 308 Faculté de Médecine, Université de Nancy, Nancy, France
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Jean-Louis GUEANT
Jean-Louis GUEANT
§
*Laboratoire de Biologie Cellulaire et Moléculaire en Nutrition et INSERM Unité 308 Faculté de Médecine, Université de Nancy, Nancy, France
§To whom correspondence should be addressed.
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Publisher: Portland Press Ltd
Received:
June 30 1995
Revision Received:
September 04 1995
Accepted:
September 12 1995
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1996
1996
Biochem J (1996) 313 (2): 675–681.
Article history
Received:
June 30 1995
Revision Received:
September 04 1995
Accepted:
September 12 1995
Citation
Alibada YERIMA, Amal SAFI, Isabelle GASTIN, Jean-Claude MICHALSKI, Monique SAUNIER, Jean-Louis GUEANT; Purification by cobalamin-Sepharose affinity chromatography and intrinsic factor-binding activity of an extramembrane proteolytic product from pig ileal mucosa. Biochem J 15 January 1996; 313 (2): 675–681. doi: https://doi.org/10.1042/bj3130675
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