Oxygen-regulated beta(2)-adrenergic receptor hydroxylation by EGLN3 and ubiquitylation by pVHL.
Oxygen-regulated beta(2)-adrenergic receptor hydroxylation by EGLN3 and ubiquitylation by pVHL.
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DOI：
10.1126/scisignal.2000444
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发表时间：
2009-07-07
影响因子：
7.3
通讯作者：
Stamler JS
中科院分区：
文献类型：
--
作者：
Xie L;Xiao K;Whalen EJ;Forrester MT;Freeman RS;Fong G;Gygi SP;Lefkowitz RJ;Stamler JS
激动剂诱导的异源三聚体鸟嘌呤核苷酸结合蛋白（G蛋白）偶联受体（GPCR）的泛素化和降解在表面受体稳态中起重要作用，从而调节许多生理过程。尽管β-抑制蛋白和相关的E3连接酶介导激动剂刺激的β2-肾上腺素能受体（β2AR）的溶酶体降解（一种原型GPCR），但对标记受体泛素化的分子线索和蛋白酶体对受体降解的调节仍知之甚少。我们发现，以调节缺氧诱导因子（HIF）蛋白而闻名的von Hippel-Lindau肿瘤抑制蛋白（pVHL）-E3连接酶复合物与β2AR相互作用并使其泛素化，从而降低受体丰度。我们进一步表明pVHL与β 2 AR的相互作用依赖于脯氨酸羟化（脯氨酸-382和-395），并且双加氧酶EGLN 3直接与β 2 AR相互作用，作为内源性β 2 AR脯氨酸羟化酶。在低氧条件下，受体羟基化和随后的泛素化显著降低，从而减弱受体降解和下调。值得注意的是，在细胞和组织中，内源性β2AR的丰度显示反映了EGLN 3和pVHL的组成性转换。我们的研究结果提供了深入了解GPCR调控，拓宽脯氨酰羟基化的功能范围，并扩大了我们对细胞缺氧反应的理解。
Agonist-induced ubiquitylation and degradation of heterotrimeric guanine nucleotide–binding protein (G protein)–coupled receptors (GPCRs) play an essential role in surface receptor homeostasis, thereby tuning many physiological processes. Although β-arrestin and affiliated E3 ligases mediate agonist-stimulated lysosomal degradation of the β2-adrenergic receptor (β2AR), a prototypic GPCR, the molecular cues that mark receptors for ubiquitylation and the regulation of receptor degradation by the proteasome remain poorly understood. We show that the von Hippel–Lindau tumor suppressor protein (pVHL)–E3 ligase complex, known for its regulation of hypoxia-inducible factor (HIF) proteins, interacts with and ubiquitylates the β2AR, thereby decreasing receptor abundance. We further show that the interaction of pVHL with β2AR is dependent on proline hydroxylation (proline-382 and -395) and that the dioxygenase EGLN3 interacts directly with the β2AR to serve as an endogenous β2AR prolyl hydroxylase. Under hypoxic conditions, receptor hydroxylation and subsequent ubiquitylation decrease dramatically, thus attenuating receptor degradation and down-regulation. Notably, in both cells and tissue, the abundance of endogenous β2AR is shown to reflect constitutive turnover by EGLN3 and pVHL. Our findings provide insight into GPCR regulation, broaden the functional scope of prolyl hydroxylation, and expand our understanding of the cellular response to hypoxia.
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影响因子：
56.9
作者：
Jaakkola, P;Mole, DR;Ratcliffe, PJ
通讯作者：
Ratcliffe, PJ
DOI：
10.1073/pnas.1936026100
发表时间：
2003-11-25
影响因子：
11.1
作者：
Foerster, K;Groner, F;Herzig, S
通讯作者：
Herzig, S
DOI：
10.1164/ajrccm.158.supplement_2.13tac110
发表时间：
1998-11-01
影响因子：
24.7
作者：
Johnson, M
通讯作者：
Johnson, M
影响因子：
5.3
作者：
Bishop, Tammie;Gallagher, Denis;Ratcliffe, Peter J.
通讯作者：
Ratcliffe, Peter J.
影响因子：
64.8
作者：
Hon, WC;Wilson, MI;Jones, EY
通讯作者：
Jones, EY