Some properties of a purinergic receptor solubilized from human uterus membranes

• Published in: FEBS letters Vol. 172; no. 2; pp. 335 - 338
• Main Authors: Ronca-Testoni, S., Galbani, P., Gambacciani, M.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 09.07.1984; Elsevier
• Subjects: Adenosine - analogs & derivatives; Adenosine - metabolism; Adenosine-5'-(N-ethylcarboxamide); Biological and medical sciences; Cell Membrane - metabolism; Cell receptors; Cell structures and functions; CHA, N 6-cyclohexyladenosine; Cholic Acid; Cholic Acids; Drug Stability; Female; Fundamental and applied biological sciences. Psychology; Human uterus membrane; Humans; Kinetics; L-PIA, L-N 6-phenylisopropyladenosine; man; Molecular and cellular biology; Molecular Weight; Myometrium - metabolism; NECA, 5'-N-ethylcarboxamideadenosine; NEM, N-ethylmaleimide; p-CMPS, p-chloromercuriphenylsulfonic acid; purinergic; Purinergic receptor 5'-N-Ethylcarboxamideadenosine; Purinergic receptor5'-N-Ethylcarboxamideadenosine; Receptor solubilization; Receptors, Cell Surface - metabolism; Receptors, Purinergic; Solubility; uterus; Purinergic receptor 5'-N-Ethylcarboxamideadenosine; NEM, N-ethylmaleimide; p-CMPS, p-chloromercuriphenylsulfonic acid; L-PIA, L- N 6-phenylisopropyladenosine; Human uterus membrane; CHA, N 6-cyclohexyladenosine; Receptor solubilization; NECA, 5'- N-ethylcarboxamideadenosine; Human; Adenosine; Uterus; Affinity labelling; Purinergic receptor; Woman; Solubilization
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(84)81152-7

A purinergic receptor was identified in human myometrium membranes using 5'- N-[ 3H]ethylcarboxamideadenosine ([ 3H]NECA) as radioligand. Scatchard analysis of the binding data gave a K d of 123 nM with 2.3pmol ligand bound mg protein. Displacement studies indicated that the binding site had the characteristics of the A 2 adenosine receptors and some of those of the P 2 purinoceptors since it was inhibited by two slowly degradable ATP derivatives with IC 50 values comparable to that of NECA. The receptor was solubilized with sodium cholate and its binding properties were the same as those of the membrane-bound form. No -SH group appeared to be essential for the binding activity. By density gradient centrifugation the purinergic receptor-detergent complex was estimated to have an apparent molecular mass of 95 kDa.