Glycoprotein nature of α2‐adrenergic receptors labeled with p‐azido[3H] clonidine in calf retina membranes

FEBS Letters

Convents, André¹  Vauquelin, Georges¹  Van Driessche, Edilbert¹  Beeckmans, Sonia¹  Convents, Daniel¹  De Backer, Jean-Paul¹ 

[1] Department of Protein Chemistry, Instituut voor Molekulaire Biologie, Vrije Universiteit Brussel, Brussels, Belgium

关键词: α2-Adrenergic receptor;    p-Azido[3H]clonidine;    Lectin affinity chromatography;    Glycosidase;    (Calf retina);    [3H]PAZ;    p-azido[3H]clonidine;    PMSF;    phenylmethylsulfonyl fluoride;    Con A;    concanavalin A;    WGA;    wheat germ agglutinin;    SDS-PAGE;    SDS-polyacrylamide gel electrophoresis;    TFMS;    trifluoromethanesulfonic acid;   

DOI  :  10.1016/0014-5793(88)80142-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

α₂,-Adrenergic receptors in calf retina membranes can be specifically labeled with the tritiated agonist p-azido[³H]clonidine. Saturation binding in the dark occurs with high affinity (1.3 ± 0.3 nM) to a single class of sites (1122 ± 67 fmol/mg protein). Irradiation of the membrane-bound radioligand results in the labeling of a peptide band with an apparent size of 65 kDa and a characteristic pharmacological profile for an α₂-adrenergic receptor. The carbohydrate moieties of the α₂-receptor are characterized by lectin affinity chromatography and glycosidase treatment. The Nonidet P-40-solubilized, p-azido[³H]clonidine-labeled receptors are completely retained by Con A- as well as WGA-Sepharose columns. Neuraminidase, α-mannosidase and TFMS do not affect the electrophoretic mobility of the receptor on SDS-PAGE whereas endoglycosidase F reduces the apparent size to 45 kDa.

【 授权许可】

Unknown