Characterization of human and rodent native and recombinant adenosine A(2B) receptors by radioligand binding studies.
Characterization of human and rodent native and recombinant adenosine A(2B) receptors by radioligand binding studies.
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DOI：
10.1007/s11302-006-9012-4
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发表时间：
2006-09
影响因子：
3.5
通讯作者：
Muller, Christa E
中科院分区：
文献类型：
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作者：
Bertarelli, Daniela C G;Diekmann, Martina;Hayallah, Alaa M;Rusing, Dorothee;Iqbal, Jamshed;Preiss, Birgit;Verspohl, Eugen J;Muller, Christa E
在放射性配体结合研究中，使用[3 H]PSB-298 [（8-{4-[2-（2-羟乙基氨基）-2-氧代乙氧基]苯基}-1-丙基黄嘌呤]研究了天然人和啮齿动物细胞系的腺苷A2 B受体。[~ 3 H]PSB-298具有饱和可逆结合。它对人胚肾细胞（HEK-293）中表达的重组人腺苷A2 B受体表现出60 ± 1 nM的KD值和有限的能力（Bmax = 3.511 fmol/mg蛋白）。氯化钠（100 mM）的加入导致放射性配体识别的结合位点的数量增加了三倍。5′-N-乙基甲酰氨基腺苷（NECA）在NaCl存在下的曲线右移，而拮抗剂PSB-298的曲线左移，表明PSB-298可能是A2 B受体的反向激动剂。腺苷A2 B受体是小鼠神经母细胞瘤×大鼠神经胶质瘤杂交细胞系NG 108 -15细胞上的主要腺苷A2受体亚型。在大鼠INS-1细胞（胰岛素分泌细胞系）中的结合研究表明，[3 H]PSB-298是该细胞系中腺苷A2 B结合位点的选择性放射性配体。
Adenosine A2B receptors of native human and rodent cell lines were investigated using [3H]PSB-298 [(8-{4-[2-(2-hydroxyethylamino)-2-oxoethoxy]phenyl}-1-propylxanthine] in radioligand binding studies. [3H]PSB-298 showed saturable and reversible binding. It exhibited a KD value of 60 ± 1 nM and limited capacity (Bmax = 3.511 fmol per milligram protein) at recombinant human adenosine A2B receptors expressed in human embryonic kidney cells (HEK-293). The addition of sodium chloride (100 mM) led to a threefold increase in the number of binding sites recognized by the radioligand. The curve of the agonist 5′-N-ethylcarboxamidoadenosine (NECA) was shifted to the right in the presence of NaCl, while the curve of the antagonist PSB-298 was shifted to the left, indicating that PSB-298 may be an inverse agonist at A2B receptors. Adenosine A2B receptors were shown to be the major adenosine A2 receptor subtype on the mouse neuroblastoma x rat glioma hybrid cell line NG108-15 cells. Binding studies at rat INS-1 cells (insulin secreting cell line) demonstrated that [3H]PSB-298 is a selective radioligand for adenosine A2B binding sites in this cell line.