Catecholamine binding to plasma membrane enriched fractions of heart and skeletal muscle.
Binding of [3H]epinephrine to plasma membrane enriched fractions from guinea pig heart and rabbit skeletal muscle was investigated using the micropore filtration technique. [3H]Epinephrine and [3H]norepinephrine were found to be degraded rapidly in aqueous buffer at pH 7.6 and 37 degrees C. Deterioration of the compounds could be prevented by low concentrations of dithiothreitol. Binding of [3H]epinephrine to both membrane preparations was a slow process requiring 60 min to approach equilibrium in the case of cardiac membranes at 37 degrees C, and 20 min for skeletal muscle membranes at O degrees C. Binding was antagonized by the unlabeled beta-agonists, isopropylnorepinephrine, epinephrine, and norepinephrine but all were equipotent. A variety of catechol compounds were as effective antagonists of binding as the catecholamines. The beta-adrenergic antagonists propranolol, pronethalol, and dichloroisoproterenol were not effective in inhibiting binding to either membrane preparation. D-Norepinephrine and L-norepinephrine were equi-effective in antagonizing binding of [3H]norephinephrine to skeletal muscle membranes. It was concluded that binding of labeled catecholamine to isolated tissue membranes using the micropore filtration technique does not represent interaction with the specific beta-adrenergic receptor, but more likely reflects a less specific binding of compounds having one or more hydroxyl groups on a ring.
D000318:Adrenergic beta-Agonists / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002395:Catecholamines / Q000032:analysis / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002855:Chromatography, Thin Layer; D004229:Dithiothreitol; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008297:Male; D008851:Micropore Filters; D009132:Muscles / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism*; D011485:Protein Binding; D011817:Rabbits; D011941:Receptors, Adrenergic; D012508:Sarcolemma / Q000378:metabolism; D013053:Spectrophotometry; D014316:Tritium; D014462:Ultrafiltration / Q000379:methods
D000318:Adrenergic beta-Agonists; D002395:Catecholamines; D011941:Receptors, Adrenergic; D014316:Tritium; D004229:Dithiothreitol