Tags

Type your tag names separated by a space and hit enter

Binding assay and physicochemical characteristics of solubilized intrinsic factor receptor in ileal mucosal homogenates using phenyl-Sepharose to separate the saturated receptor from free intrinsic factor.
Biochim Biophys Acta. 1991 Apr 09; 1073(3):614-8.BB

Abstract

A radioisotopic assay was set to determine the physicochemical properties of the solubilized intrinsic factor receptor in pig mucosal extracts. In this assay, phenyl-Sepharose was used to separate the receptor-intrinsic factor-labelled cobalamin complex from the free saturated intrinsic factor. The association constant (at pH 7.4) of the receptor-intrinsic factor complex was estimated at 3.4 +/- 0.3 nM-1. Adsorption of the apo-receptor to phenyl-Sepharose allowed its binding site to be made accessible to intrinsic factor with an association constant in order of 6 nM-1. The receptor binding activity obtained with five mucosal extracts was closely correlated with that obtained by gel filtration of the intrinsic factor-receptor complex (r = 0.99). The radioisotope assay was used to detect the unsaturated receptor (apo-receptor) in sucrose density ultracentrifugation and in superose 6 gel filtration. The sedimentation coefficient was 9.5 s. The apo-receptor was eluted in three peaks in gel filtration, corresponding to the formation of oligomers. The peak of the monomer was increased in presence of EDTA. Its molecular mass was estimated at 270 kDa and its Stokes radius at 5.9 nm. It was concluded that calcium is involved in the oligomerisation of the apo-receptor.

Authors+Show Affiliations

Group of Biochemistry-Immunology, INSERM U 308 University of Nancy I, Vandoeuvre-lès-Nancy, France.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

1849746

Citation

Guéant, J L., et al. "Binding Assay and Physicochemical Characteristics of Solubilized Intrinsic Factor Receptor in Ileal Mucosal Homogenates Using phenyl-Sepharose to Separate the Saturated Receptor From Free Intrinsic Factor." Biochimica Et Biophysica Acta, vol. 1073, no. 3, 1991, pp. 614-8.
Guéant JL, Schohn H, Brulé H, et al. Binding assay and physicochemical characteristics of solubilized intrinsic factor receptor in ileal mucosal homogenates using phenyl-Sepharose to separate the saturated receptor from free intrinsic factor. Biochim Biophys Acta. 1991;1073(3):614-8.
Guéant, J. L., Schohn, H., Brulé, H., Saunier, M., Gräsbeck, R., & Nicolas, J. P. (1991). Binding assay and physicochemical characteristics of solubilized intrinsic factor receptor in ileal mucosal homogenates using phenyl-Sepharose to separate the saturated receptor from free intrinsic factor. Biochimica Et Biophysica Acta, 1073(3), 614-8.
Guéant JL, et al. Binding Assay and Physicochemical Characteristics of Solubilized Intrinsic Factor Receptor in Ileal Mucosal Homogenates Using phenyl-Sepharose to Separate the Saturated Receptor From Free Intrinsic Factor. Biochim Biophys Acta. 1991 Apr 9;1073(3):614-8. PubMed PMID: 1849746.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Binding assay and physicochemical characteristics of solubilized intrinsic factor receptor in ileal mucosal homogenates using phenyl-Sepharose to separate the saturated receptor from free intrinsic factor. AU - Guéant,J L, AU - Schohn,H, AU - Brulé,H, AU - Saunier,M, AU - Gräsbeck,R, AU - Nicolas,J P, PY - 1991/4/9/pubmed PY - 1991/4/9/medline PY - 1991/4/9/entrez SP - 614 EP - 8 JF - Biochimica et biophysica acta JO - Biochim Biophys Acta VL - 1073 IS - 3 N2 - A radioisotopic assay was set to determine the physicochemical properties of the solubilized intrinsic factor receptor in pig mucosal extracts. In this assay, phenyl-Sepharose was used to separate the receptor-intrinsic factor-labelled cobalamin complex from the free saturated intrinsic factor. The association constant (at pH 7.4) of the receptor-intrinsic factor complex was estimated at 3.4 +/- 0.3 nM-1. Adsorption of the apo-receptor to phenyl-Sepharose allowed its binding site to be made accessible to intrinsic factor with an association constant in order of 6 nM-1. The receptor binding activity obtained with five mucosal extracts was closely correlated with that obtained by gel filtration of the intrinsic factor-receptor complex (r = 0.99). The radioisotope assay was used to detect the unsaturated receptor (apo-receptor) in sucrose density ultracentrifugation and in superose 6 gel filtration. The sedimentation coefficient was 9.5 s. The apo-receptor was eluted in three peaks in gel filtration, corresponding to the formation of oligomers. The peak of the monomer was increased in presence of EDTA. Its molecular mass was estimated at 270 kDa and its Stokes radius at 5.9 nm. It was concluded that calcium is involved in the oligomerisation of the apo-receptor. SN - 0006-3002 UR - https://www.unboundmedicine.com/medline/citation/1849746/Binding_assay_and_physicochemical_characteristics_of_solubilized_intrinsic_factor_receptor_in_ileal_mucosal_homogenates_using_phenyl_Sepharose_to_separate_the_saturated_receptor_from_free_intrinsic_factor_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0304-4165(91)90238-C DB - PRIME DP - Unbound Medicine ER -