Purification of the photoaffinity-labeled glucagon receptor by gel electrophoretic methods.
Rat liver plasma membrane glucagon receptor has been purified with a yield of 0.01% to an estimated homogeneity of 32-60%, using a 2-stage electrophoretic procedure. SDS-solubilized membrane proteins labeled by the photoaffinity-agent, Ne-4-azidophenylamidinoglucagon (APA-glucagon), were separated by polyacrylamide gel electrophoresis in SDS-containing buffers. Gel slices corresponding to the molecular weight of the receptor were excised, electrophoretically extracted and concentrated. The concentrate was subjected to isoelectric focusing on Sephadex to yield a purified product in which the photoaffinity-labeled receptor, with a molecular weight of 56K and a pI' of 5.9, is the sole major component.
D000345:Affinity Labels; D000818:Animals; D001386:Azides; D002462:Cell Membrane / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005934:Glucagon / Q000031:analogs & derivatives* / Q000378:metabolism; D007525:Isoelectric Focusing; D008099:Liver / Q000378:metabolism*; D008970:Molecular Weight; D051381:Rats; D011956:Receptors, Cell Surface / Q000302:isolation & purification* / Q000378:metabolism; D018027:Receptors, Glucagon
D000345:Affinity Labels; D001386:Azides; D011956:Receptors, Cell Surface; D018027:Receptors, Glucagon; C041462:glucagon, 12-(N(6)-(4-azidophenylamidino)Lys)-; D005934:Glucagon