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Solubilization of the ileal receptor for intrinsic factor--vitamin B-12 complex by digestion with papain.
Biochem J. 1981 Nov 15; 200(2):225-9.BJ

Abstract

Brush-border-membrane vesicles isolated from hamster ileum were incubated with either papain or Pronase P and subsequently centrifuged to obtain soluble (supernatant) and insoluble (pellet) fractions. Papain (4 units/ml) solubilized 95--100% of the sucrase and leucine naphthylamide-hydrolysing activities but only 30% of the alkaline phosphatase. Digestion with papain also resulted in the solubilization of more than 75% of the ileal receptor for intrinsic factor-vitamin B-12 complex with a corresponding decrease in receptor activity in the pellet. Essentially 100% of the receptor activity was recovered. In contrast, digestion with Pronase P resulted in a decrease in total receptor activity. Papain-solubilized receptor was not sedimented by centrifugation at 105 000 g for 90 min and was eluted in the included volume of Sepharose 6B. Like the binding to more intact preparations, binding of intrinsic factor-vitamin B-12 complex to papain-solubilized receptor was rapid, reaching 50% of maximum in 8 min, and required Ca2+. Although Mg2+ could not completely substitute for Ca2+, Mg2+ did stimulate Ca2+-dependent binding at low Ca2+ concentrations. These results demonstrate that the ileal receptor for intrinsic factor-vitamin B-12 complex can be solubilized with papain, and suggest that papain solubilization may be a useful first step in the isolation and purification of this receptor.

Authors

No affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

6280680

Citation

Beesley, R C.. "Solubilization of the Ileal Receptor for Intrinsic Factor--vitamin B-12 Complex By Digestion With Papain." The Biochemical Journal, vol. 200, no. 2, 1981, pp. 225-9.
Beesley RC. Solubilization of the ileal receptor for intrinsic factor--vitamin B-12 complex by digestion with papain. Biochem J. 1981;200(2):225-9.
Beesley, R. C. (1981). Solubilization of the ileal receptor for intrinsic factor--vitamin B-12 complex by digestion with papain. The Biochemical Journal, 200(2), 225-9.
Beesley RC. Solubilization of the Ileal Receptor for Intrinsic Factor--vitamin B-12 Complex By Digestion With Papain. Biochem J. 1981 Nov 15;200(2):225-9. PubMed PMID: 6280680.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Solubilization of the ileal receptor for intrinsic factor--vitamin B-12 complex by digestion with papain. A1 - Beesley,R C, PY - 1981/11/15/pubmed PY - 1981/11/15/medline PY - 1981/11/15/entrez SP - 225 EP - 9 JF - The Biochemical journal JO - Biochem J VL - 200 IS - 2 N2 - Brush-border-membrane vesicles isolated from hamster ileum were incubated with either papain or Pronase P and subsequently centrifuged to obtain soluble (supernatant) and insoluble (pellet) fractions. Papain (4 units/ml) solubilized 95--100% of the sucrase and leucine naphthylamide-hydrolysing activities but only 30% of the alkaline phosphatase. Digestion with papain also resulted in the solubilization of more than 75% of the ileal receptor for intrinsic factor-vitamin B-12 complex with a corresponding decrease in receptor activity in the pellet. Essentially 100% of the receptor activity was recovered. In contrast, digestion with Pronase P resulted in a decrease in total receptor activity. Papain-solubilized receptor was not sedimented by centrifugation at 105 000 g for 90 min and was eluted in the included volume of Sepharose 6B. Like the binding to more intact preparations, binding of intrinsic factor-vitamin B-12 complex to papain-solubilized receptor was rapid, reaching 50% of maximum in 8 min, and required Ca2+. Although Mg2+ could not completely substitute for Ca2+, Mg2+ did stimulate Ca2+-dependent binding at low Ca2+ concentrations. These results demonstrate that the ileal receptor for intrinsic factor-vitamin B-12 complex can be solubilized with papain, and suggest that papain solubilization may be a useful first step in the isolation and purification of this receptor. SN - 0264-6021 UR - https://www.unboundmedicine.com/medline/citation/6280680/Solubilization_of_the_ileal_receptor_for_intrinsic_factor__vitamin_B_12_complex_by_digestion_with_papain_ L2 - https://portlandpress.com/biochemj/article-lookup/doi/10.1042/bj2000225 DB - PRIME DP - Unbound Medicine ER -