Soluble serotonin and catecholamine binding proteins in the bovine adrenal medulla.
The soluble serotonin-binding proteins (SBP) present in the adrenal medulla and in chromaffin cells, are very similar to those reported for the bovine brain and retina. Binding of [3H]serotonin and [3H]dopamine to these SBP is increased by Fe2+ but not by Fe3+. At an optimal concentration of Fe2+ (0.1 mM) these proteins behave as a single class of non-cooperative sites for [3H]serotonin (Bmax = 124 +/- 28 pmol/mg protein, KD = 0.51 +/- 0.13 microM) and [3H]dopamine (Bmax = 685 +/- 118 pmol/mg protein, KD = 0.46 +/- 0.06 microM). Binding of [3H]dopamine is also increased by Cu2+ and Mn2+, but to a lesser extent than by Fe2+. Catecholamines are good competitors for [3H]serotonin binding (Ki = 0.31 microM for dopamine, 0.6 microM for adrenaline and 0.9 microM for noradrenaline). The serotonin binding proteins from adrenal medulla elute in the void volume of a Sephacryl 100 HR gel filtration column, reflecting aggregation, and migrate mainly with an apparent molecular weight of 45 kDa in native polyacrylamide gel electrophoresis experiments. Subcellular localization studies and release experiments suggest that SBP are not present in chromaffin granules, but in the cytosol of purified chromaffin cells. The present data suggest that these proteins must have other functions than storing monoamines in synaptic vesicles.
Pinxteren
J
J
Department of Neuropharmacology, Universitaire Instelling Antwerpen, Wilrijk, Belgium.
Jimenez del Rio
M
M
Velez Pardo
C
C
Ebinger
G
G
Vauquelin
G
G
De Potter
W
W
D000313:Adrenal Medulla / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000378:metabolism; D002837:Chromaffin Granules / Q000378:metabolism; D053379:Chromogranin A; D002864:Chromogranins / Q000302:isolation & purification / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004298:Dopamine / Q000378:metabolism*; D050483:Dopamine Plasma Membrane Transport Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D009638:Norepinephrine / Q000378:metabolism; D012160:Retina / Q000378:metabolism; D012701:Serotonin / Q000378:metabolism*
D002352:Carrier Proteins; D053379:Chromogranin A; D002864:Chromogranins; D050483:Dopamine Plasma Membrane Transport Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; C025366:serotonin-binding protein; D012701:Serotonin; D004298:Dopamine; D009638:Norepinephrine