Partial purification and biochemical characterization of a membrane glucocorticoid receptor from an amphibian brain.
A membrane receptor for corticosterone (mGR) in the brain of the roughskin newt (Taricha granulosa) has been previously identified. This manuscript reports the evaluation of several chromatographic resins for enrichment of the newt mGR solubilized from neuronal membranes. A protein with an apparent molecular weight of 63 kDa was purified to near homogeneity following sequential purification using ammonium sulfate fractionation, wheat germ agglutinin (WGA)-agarose chromatography, hydroxylapatite chromatography, and an immobilized ligand affinity resin (Corticosterone-Sepharose). Other studies employed a novel protein differential display strategy and a photoaffinity labeling strategy to visualize candidate receptor proteins following SDS-PAGE. Both of these techniques also identified a 63 kDa protein, agreeing with the estimation of molecular weight from the purification data. Furthermore, the use of 2D SDS-PAGE following the photolabeling procedure showed the candidate 63 kDa protein to have a pI of approximately 5.0. Taken together these data suggest that the newt mGR is an acidic glycoprotein with an apparent molecular weight of 63 kDa. Because these characteristics of newt mGR are inconsistent with the characteristics of intracellular glucocorticoid receptors, these two receptor proteins are apparently distinct.
Evans
S J
SJ
Zoology Department, Oregan State University, Corvallis, OR 97331-2914, USA.
Murray
T F
TF
Moore
F L
FL
D000645:Ammonium Sulfate / Q000737:chemistry; D000818:Animals; D001921:Brain / Q000378:metabolism*; D001923:Brain Chemistry; D002846:Chromatography, Affinity; D002847:Chromatography, Agarose / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods; D003345:Corticosterone / Q000378:metabolism; D017886:Durapatite / Q000737:chemistry; D037102:Lectins / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D019920:Photoaffinity Labels / Q000737:chemistry; D011965:Receptors, Glucocorticoid / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D012455:Salamandridae; D014909:Wheat Germ Agglutinins / Q000378:metabolism
D037102:Lectins; D008565:Membrane Proteins; D019920:Photoaffinity Labels; D011965:Receptors, Glucocorticoid; D014909:Wheat Germ Agglutinins; D017886:Durapatite; D000645:Ammonium Sulfate; D003345:Corticosterone