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Purification and Characterization of B 2 Bradykinin Receptor from Rat Uterus
Author(s) -
Yaqoob Mohammed,
Snell Christopher R.
Publication year - 1994
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1994.62010017.x
Subject(s) - bradykinin , uterus , receptor , chemistry , bradykinin receptor , endocrinology , medicine , biology , biochemistry
A B 2 bradykinin (BK) receptor was solubilised and partially purified from rat uterine membranes by a combination of ammonium sulphate precipitation, desalting on Sephadex G‐50, and hydroxyapatite and wheat germ agglutinin affinity chromatography. The partially purified BK receptor, enriched 1,500‐fold, was then cross‐linked to 125 l‐Tyr 0 ‐BK using disuccinimidyl suberate and purified to homogeneity as a single protein species on two‐dimensional gel electrophoresis with a molecular mass of 81 kDa. This molecular size was in agreement with the value of 80–120 kDa estimated from Sephacryl 300 size exclusion column chromatography of the B 2 receptor. The partially purified and the crude solubilised B 2 BK receptor from rat uterus showed similar affinities for BK and the BK analogues iodo‐Tyr 0 ‐BK, D‐Phe 7 ‐BK, and des‐Arg 9 ‐BK, indicating that the ligand binding specificity of the receptor had been retained during the purification procedures. The biochemical properties of the solubilised B 2 BK receptor correspond to those of a hydrophobic acidic glycoprotein (isoelectric focusing gave a value of 4.5–4.7) that binds specifically to wheat germ agglutinin but has no affinity for either concanavalin A or lentil lectin, suggesting the absence of terminal mannose or glucose residues.