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In the present study we have determined the molecular mass of the beta-cell sulfonylurea receptor in its native form by two different experimental approaches; gel filtration chromatography and radiation inactivation analysis. We first confirmed that the denatured photolabelled MIN6 beta-cell receptor had a molecular size of 141 +/- 2 kDa (mean +/- S.E., n = 8). Under non-denaturing conditions, using gel filtration chromatography, apparent molecular masses of 166 +/- 1 kDa (mean +/- S.E., n = 3) and 182 +/- 5 kDa (mean +/- S.E., n = 4) were determined for the photoaffinity-labelled and unlabelled sulfonylurea receptor, respectively. We conclude that in the solubilized state the receptor exists as a monomer. Radiation inactivation analysis indicated that the receptor has a target size of 250 +/- 30 kDa (mean +/- S.E., n = 7). This value for the molecular mass is larger than that obtained from SDS-PAGE following photolabelling of the receptor (141 kDa) suggesting that the beta-cell sulfonylurea receptor is composed of more than one subunit in the native membrane.

Type

Journal article

Journal

FEBS Lett

Publication Date

24/01/1994

Volume

338

Pages

98 - 102

Keywords

ATP-Binding Cassette Transporters, Affinity Labels, Cell Membrane, Cells, Cultured, Chromatography, Gel, Electrophoresis, Polyacrylamide Gel, Islets of Langerhans, Molecular Weight, Potassium Channels, Potassium Channels, Inwardly Rectifying, Receptors, Drug, Sulfonylurea Receptors