MARCH2 promotes endocytosis and lysosomal sorting of carvedilol-bound β(2)-adrenergic receptors.
MARCH2 promotes endocytosis and lysosomal sorting of carvedilol-bound β(2)-adrenergic receptors.
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DOI：
10.1083/jcb.201208192
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发表时间：
2012-11-26
期刊：
影响因子：
--
通讯作者：
Shenoy SK
中科院分区：
文献类型：
--
作者：
Han SO;Xiao K;Kim J;Wu JH;Wisler JW;Nakamura N;Freedman NJ;Shenoy SK
β2-肾上腺素能受体拮抗剂卡维地洛招募MARCH2，一种独特的E3泛素连接酶，促进受体内吞和溶酶体运输。泛素化β2肾上腺素能受体(β2ARs)的溶酶体降解是激动剂刺激引起的长期脱敏的主要机制。令人惊讶的是，β受体拮抗剂卡维地洛还诱导血管平滑肌细胞内源性表达的β2AR和HEK293细胞中过表达的FLAG-β2ARs的泛素化和溶酶体转运。卡维地洛抑制β-2AR循环，抑制Nedd4E3连接酶的募集，促进去泛素酶USP20和USP33的解离。利用蛋白质组学方法(液-质联用)，我们鉴定了E3连接酶MARCH2与卡维地洛结合的β2AR相互作用。卡维地洛可稳定MARCH2与内化β2ARs的结合，而不涉及β-arrestin。小干扰RNA介导的MARCH2抑制卡维地洛诱导的血管平滑肌细胞泛素化、内吞和内源性β2ARs的降解。这些发现强烈表明，特定的配体招募不同的E3连接酶机制来激活细胞表面受体，并指导它们的细胞内行程。在卡维地洛的β阻滞剂治疗下，MARCH2E3连接酶活性调节细胞表面β2AR的表达，从而调节其信号转导。
The β2-adrenergic receptor antagonist carvedilol recruits MARCH2, a unique E3 ubiquitin ligase, to promote receptor endocytosis and lysosomal trafficking. Lysosomal degradation of ubiquitinated β2-adrenergic receptors (β2ARs) serves as a major mechanism of long-term desensitization in response to prolonged agonist stimulation. Surprisingly, the βAR antagonist carvedilol also induced ubiquitination and lysosomal trafficking of both endogenously expressed β2ARs in vascular smooth muscle cells (VSMCs) and overexpressed Flag-β2ARs in HEK-293 cells. Carvedilol prevented β2AR recycling, blocked recruitment of Nedd4 E3 ligase, and promoted the dissociation of the deubiquitinases USP20 and USP33. Using proteomics approaches (liquid chromatography–tandem mass spectrometry), we identified that the E3 ligase MARCH2 interacted with carvedilol-bound β2AR. The association of MARCH2 with internalized β2ARs was stabilized by carvedilol and did not involve β-arrestin. Small interfering RNA–mediated down-regulation of MARCH2 ablated carvedilol-induced ubiquitination, endocytosis, and degradation of endogenous β2ARs in VSMCs. These findings strongly suggest that specific ligands recruit distinct E3 ligase machineries to activated cell surface receptors and direct their intracellular itinerary. In response to β blocker therapy with carvedilol, MARCH2 E3 ligase activity regulates cell surface β2AR expression and, consequently, its signaling.
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影响因子：
3.3
作者：
Eyster CA;Cole NB;Petersen S;Viswanathan K;Früh K;Donaldson JG
通讯作者：
Donaldson JG
影响因子：
6
作者：
Domanski, MJ;Krause-Steinrauf, H;Eichhorn, E
通讯作者：
Eichhorn, E
影响因子：
24
作者：
Jabbour, Andrew;Macdonald, Peter S.;Hayward, Christopher S.
通讯作者：
Hayward, Christopher S.
影响因子：
4.8
作者：
Ahn, S;Maudsley, S;Daaka, Y
通讯作者：
Daaka, Y
DOI：
10.1073/pnas.0804745105
发表时间：
2008-09-23
影响因子：
11.1
作者：
Kim, Il-Man;Tilley, Douglas G.;Rockman, Howard A.
通讯作者：
Rockman, Howard A.