Key amino acid residues in the melanocortin-4 receptor for nonpeptide THIQ specific binding and signaling.
Key amino acid residues in the melanocortin-4 receptor for nonpeptide THIQ specific binding and signaling.
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DOI：
10.1016/j.regpep.2009.03.006
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发表时间：
2009-06-05
影响因子：
--
通讯作者：
Harmon, Carroll M.
中科院分区：
文献类型：
--
作者：
Yang, Yingkui;Cai, Minying;Chen, Min;Qu, Hongchang;McPherson, David;Hruby, Victor;Harmon, Carroll M.
黑皮质素 4 受体 (MC4R) 在食物摄入和葡萄糖稳态的调节中发挥着重要作用。合成非肽化合物 N- (3R)-1 4-四氢异喹啉鎓-3-基羰基 -(1R)-1-(4-氯苄基)-2- 4-环己基-4-(1H-1,2,4-三唑-1-基甲基)哌啶-1-基 -2-氧代乙胺 (THIQ) 是一种有效的 激动 MC4R 但不激动 hMC2R。在这项研究中，我们利用两种方法（嵌合受体和定点诱变）来缩小负责 THIQ 结合和信号传导的 MC4R 关键氨基酸残基范围。构建了用 hMC2R 同源区替换人 MC4R (hMC4R) 的第二、第三、第四、第五和第六跨膜区 (TM) 的盒。我们的结果表明，除了 hMC4R TM3 的替换之外，hMC4R 的这些 TM 与 hMC2R 同源区域的盒替换并未显着改变 THIQ 结合亲和力和效力，这表明 hMC4R 的这些 TM 中的保守氨基酸残基是 THIQ 结合和信号传导的主要潜在候选者，而 MC4R 的 TM3 中的非保守残基可能会影响 THIQ 的结合亲和力和效力。 也参与其中。然后创建了 19 个 MC4R 突变体，包括 13 个保守氨基酸残基和 6 个非保守氨基酸残基。我们的结果表明，七个保守残基 [E100 (TM2)、D122 (TM3)、D126 (TM3)、F254 (TM6)、W258 (TM6)、F261 (TM6)、H264 (TM6)] 对于 THIQ 结合很重要，三个非保守残基 [N123 (TM3)、I129 (TM3) 和 S131 (TM3)] 参与 THIQ 选择性。总之，我们的结果表明 THIQ 利用保守和非保守氨基酸残基在 hMC4R 上进行结合和信号传导，非保守残基可能负责 MC4R 选择性。
Melanocortin 4 receptor (MC4R) plays an important role in the regulation of food intake and glucose homeostasis. Synthetic nonpeptide compound N- (3R)-1 4-tetrahydroisoquinolinium-3-ylcarbonyl -(1R)-1-(4-chlorobenzyl)-2- 4-cyclohexyl-4-(1H-1,2,4-triazol-1-ylmethyl)piperidin-1-yl -2-oxoethylamine (THIQ) is a potent agonist at MC4R but not at hMC2R. In this study, we utilized two approaches (chimeric receptor and site-directed mutagenesis) to narrow down the key amino acid residues of MC4R responsible for THIQ binding and signaling. Cassette substitutions of the second, third, fourth, fifth, and sixth transmembrane regions (TMs) of the human MC4R (hMC4R) with the homologous regions of hMC2R were constructed. Our results indicate that the cassette substitutions of these TMs of the hMC4R with homologous regions of the hMC2R did not significantly alter THIQ binding affinity and potency except the substitution of the hMC4R TM3, suggesting that the conserved amino acid residues in these TMs of the hMC4R are main potential candidates for THIQ binding and signaling while non conserved residues in TM3 of MC4R may also be involved. Nineteen MC4R mutants were then created, including 13 conserved amino acid residues and 6 non-conserved amino acid residues. Our results indicate that seven conserved residue [E100 (TM2), D122 (TM3), D126 (TM3), F254 (TM6), W258 (TM6), F261 (TM6), H264 (TM6)] are important for THIQ binding and three non-conserved residues [N123 (TM3), I129 (TM3) and S131 (TM3)] are involved in THIQ selectivity. In conclusion, our results suggest that THIQ utilize both conserved and non-conserved amino acid residues for binding and signaling at hMC4R and non conserved residues may be responsible for MC4R selectivity.
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影响因子：
2.9
作者：
Chen, Min;Aprahamian, Charles J.;Yang, Yingkui
通讯作者：
Yang, Yingkui
影响因子：
7.3
作者：
Chen, C;Pontillo, J;Saunders, J
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Saunders, J
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4
作者：
Conrad, DM;Hanniman, EA;Hoskin, DW
通讯作者：
Hoskin, DW
影响因子：
11.4
作者：
BALDWIN, JM
通讯作者：
BALDWIN, JM
影响因子：
64.8
作者：
Fan, W;Boston, BA;Cone, RD
通讯作者：
Cone, RD