Similar ligand binding in recombinant human alpha 2 C2-adrenoceptors produced in mammalian, insect and yeast cells.
Ligand binding properties were investigated in recombinant human alpha 2C2-adrenoceptors expressed in three different host systems: Shionogi S115 mouse mammary tumour cells, Spodoptera frugiperda Sf9 insect cells and Saccharomyces cerevisiae yeast cells. The expected 43 kDa alpha 2C2 protein was visualized with immunoblotting using a polyclonal alpha 2C2-receptor antibody. [3H]Rauwolscine binding in cell homogenates or membranes (Bmax 3-11 pmol/mg protein; Kd approximately 5.5 nM) was inhibited by prazosin, oxymetazoline, RX821002, chlorpromazine and (-)-noradrenaline with and without the GTP-analogue Gpp(NH)p with similar Ki values in the different host systems. This indicates that alpha 2C2-adrenoceptors retain their binding characteristics irrespective of the host environment.
Marjamäki
A
A
Department of Pharmacology, University of Turku, Finland.
Pohjanoksa
K
K
Ala-Uotila
S
S
Sizmann
D
D
Oker-Blom
C
C
Kurose
H
H
Scheinin
M
M
D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D002746:Chlorpromazine / Q000494:pharmacology; D004146:Dioxanes / Q000494:pharmacology; D006801:Humans; D019329:Idazoxan / Q000031:analogs & derivatives; D015151:Immunoblotting; D008325:Mammary Neoplasms, Experimental / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D009036:Moths / Q000378:metabolism*; D010109:Oxymetazoline / Q000494:pharmacology; D011224:Prazosin / Q000494:pharmacology; D011942:Receptors, Adrenergic, alpha / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured; D015016:Yohimbine / Q000378:metabolism*
D000317:Adrenergic alpha-Antagonists; D004146:Dioxanes; D011942:Receptors, Adrenergic, alpha; D011994:Recombinant Proteins; D015016:Yohimbine; D010109:Oxymetazoline; C058525:2-methoxyidazoxan; D002746:Chlorpromazine; D011224:Prazosin; D019329:Idazoxan