title: Characterization of a novel iodinated ligand, IPMPP, for human dopamine D4 receptors expressed in CHO cells.
abstract: A novel radioiodinated ligand with a high specific activity (2,200 Ci/mmol), 3-[4-(4-iodophenyl)piperazin-1-yl]methyl-1H-pyrrolo(2,3-b)pyridine ([125I]IPMPP), was successfully prepared. Binding characteristics of [125I]IPMPP were evaluated using human dopamine D4 (D4.2 variant) receptors expressed in Chinese hamster ovary (CHO) cells. Saturation analysis revealed high-affinity binding sites for [125I]IPMPP (Kd = 0.39 +/- 0.18 nM). The number of D4 receptors labeled with [125I]IPMPP at room temperature was four times higher than that labeled with [125I]S(-)5-OH-PIPAT, a radioiodinated agonist ligand (572 fmol/mg protein vs. 125 fmol/mg protein). A significant decrease in the number of binding sites was observed with [125I]S(-)5-OH-PIPAT when assays were carried out at a higher temperature (37 degrees C vs. 25 degrees C). In contrast to [125I]S(-)5-OH-PIPAT, [125I]IPMPP labeled more D4 sites at 37 degrees C. Neither magnesium ion nor guanylimidodiphosphate (Gpp(NH)p) affected [125I]IPMPP binding. These data support the conclusion that [125I]IPMPP is an antagonist ligand. The potency of various compounds, including clozapine, to inhibit [125I]IPMPP binding is consistent with the rank order measured with other radioligands for D4 receptors. In addition, measuring D4 receptor stimulation of [35S]GTPgammaS binding further demonstrated the antagonist property of IPMPP.
authors.affiliation: Department of Radiology, University of Pennsylvania, Philadelphia 19104, USA. kungmp@sunmac.spect.upenn.edu
authors.forename: M P
authors.initials: MP
authors.lastname: Kung
authors.forename: D A
authors.initials: DA
authors.lastname: Stevenson
authors.forename: Z P
authors.initials: ZP
authors.lastname: Zhuang
authors.forename: J M
authors.initials: JM
authors.lastname: Vessotskie
authors.forename: S
authors.initials: S
authors.lastname: Chumpradit
authors.forename: X M
authors.initials: XM
authors.lastname: Sun
authors.forename: H F
authors.initials: HF
authors.lastname: Kung
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D016466:CHO Cells; D006224:Cricetinae; D018491:Dopamine Agonists / Q000378:metabolism* / Q000494:pharmacology; D006150:Guanine Nucleotides / Q000494:pharmacology; D006801:Humans; D008274:Magnesium / Q000494:pharmacology; D011725:Pyridines / Q000378:metabolism* / Q000494:pharmacology; D011758:Pyrroles / Q000378:metabolism* / Q000494:pharmacology; D011869:Radioligand Assay; D017448:Receptors, Dopamine D2 / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D050638:Receptors, Dopamine D4; D011994:Recombinant Proteins / Q000819:agonists / Q000235:genetics / Q000378:metabolism
chemical_list: C493599:DRD4 protein, human; D018491:Dopamine Agonists; D006150:Guanine Nucleotides; C103081:L 750667; D011725:Pyridines; D011758:Pyrroles; D017448:Receptors, Dopamine D2; D011994:Recombinant Proteins; D050638:Receptors, Dopamine D4; D008274:Magnesium