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Purification by cobalamin-Sepharose affinity chromatography and intrinsic factor-binding activity of an extramembrane proteolytic product from pig ileal mucosa.
Biochem J. 1996 Jan 15; 313 (Pt 2):675-81.BJ

Abstract

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 17,300, a yield of 63% and a cobalamin-binding activity of 11,260 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.

Authors+Show Affiliations

Laboratoire de Biologie Cellulaire et Moléculaire en Nutrition et INSERM Unité 308 Faculté de Médecine, Université de Nancy, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

8573109

Citation

Yerima, A, et al. "Purification By cobalamin-Sepharose Affinity Chromatography and Intrinsic Factor-binding Activity of an Extramembrane Proteolytic Product From Pig Ileal Mucosa." The Biochemical Journal, vol. 313 (Pt 2), 1996, pp. 675-81.
Yerima A, Safi A, Gastin I, et al. Purification by cobalamin-Sepharose affinity chromatography and intrinsic factor-binding activity of an extramembrane proteolytic product from pig ileal mucosa. Biochem J. 1996;313 (Pt 2):675-81.
Yerima, A., Safi, A., Gastin, I., Michalski, J. C., Saunier, M., & Gueant, J. L. (1996). Purification by cobalamin-Sepharose affinity chromatography and intrinsic factor-binding activity of an extramembrane proteolytic product from pig ileal mucosa. The Biochemical Journal, 313 (Pt 2), 675-81.
Yerima A, et al. Purification By cobalamin-Sepharose Affinity Chromatography and Intrinsic Factor-binding Activity of an Extramembrane Proteolytic Product From Pig Ileal Mucosa. Biochem J. 1996 Jan 15;313 (Pt 2):675-81. PubMed PMID: 8573109.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Purification by cobalamin-Sepharose affinity chromatography and intrinsic factor-binding activity of an extramembrane proteolytic product from pig ileal mucosa. AU - Yerima,A, AU - Safi,A, AU - Gastin,I, AU - Michalski,J C, AU - Saunier,M, AU - Gueant,J L, PY - 1996/1/15/pubmed PY - 1996/1/15/medline PY - 1996/1/15/entrez SP - 675 EP - 81 JF - The Biochemical journal JO - Biochem J VL - 313 (Pt 2) N2 - 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 17,300, a yield of 63% and a cobalamin-binding activity of 11,260 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. SN - 0264-6021 UR - https://www.unboundmedicine.com/medline/citation/8573109/Purification_by_cobalamin_Sepharose_affinity_chromatography_and_intrinsic_factor_binding_activity_of_an_extramembrane_proteolytic_product_from_pig_ileal_mucosa_ L2 - https://portlandpress.com/biochemj/article-lookup/doi/10.1042/bj3130675 DB - PRIME DP - Unbound Medicine ER -