Distinct Role of GRK3 in Platelet Activation by Desensitization of G Protein-Coupled Receptors.

IF 5 2区医学 Q1 HEMATOLOGY Thrombosis and haemostasis Pub Date : 2024-11-11 DOI:10.1055/a-2442-9031

Preeti K Chaudhary, Sanggu Kim, Satya P Kunapuli, Soochong Kim

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Abstract

Background: Many platelet agonists mediate their cellular effects through G protein-coupled receptors (GPCRs) to induce platelet activation, and GPCR kinases (GRKs) have been demonstrated to have crucial roles in most GPCR functions in other cell types. Here, we investigated the functional role of GRK3 and the molecular basis for the regulation of GPCR desensitization by GRK3 in platelets.

Methods: We used mice lacking GRK3 as well as β-arrestin2, which has been shown to be important in GPCR function in platelets.

Results: Platelet aggregation and dense granule secretion induced by 2-MeSADP, U46619, thrombin, and AYPGKF were significantly potentiated in both GRK3 -/- and β-arrestin2 -/- platelets compared with wild-type (WT) platelets, whereas non-GPCR agonist collagen-induced platelet aggregation and secretion were not affected. We have previously shown that GRK6 is not involved in the regulation of G_q-coupled 5HT_2A and G_z-coupled α_2A adrenergic receptors. Interestingly, in contrast to GRK6, platelet aggregation induced by costimulation of serotonin and epinephrine, which activate 5-HT_2A and α_2A adrenergic receptors, respectively, was significantly potentiated in GRK3 -/- platelets, suggesting that GRK3 is involved in general GPCR regulation. In addition, platelet aggregation in response to the second challenge of adenosine diphosphate was restored in GRK3 -/- platelets, whereas restimulation of the agonist failed to induce aggregation in WT platelets, confirming that GRK3 contributes to general GPCR desensitization. Furthermore, 2-MeSADP- and AYPGKF-induced AKT and ERK phosphorylation were significantly potentiated in GRK3 -/- platelets. Finally, GRK3 -/- mice showed shorter tail bleeding times compared with WT, indicating that GRK3 -/- mice is more susceptible to hemostasis.

Conclusion: GRK3 plays a crucial role in the regulation of platelet activation through general GPCR desensitization in platelets.

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GRK3在通过G蛋白偶联受体脱敏激活血小板过程中的独特作用

背景：许多血小板激动剂通过G蛋白偶联受体（GPCRs）介导其细胞效应以诱导血小板活化，而GPCR激酶（GRKs）已被证明在其他细胞类型的大多数GPCR功能中具有关键作用。在此，我们研究了 GRK3 的功能作用以及 GRK3 在血小板中调控 GPCR 脱敏的分子基础：方法：我们使用了缺乏 GRK3 和 β-arrestin2 的小鼠：结果：与野生型（WT）血小板相比，GRK3 -/-和 β-arrestin2 -/-血小板在 2-MeSADP、U46619、凝血酶和 AYPGKF 诱导下的血小板聚集和致密颗粒分泌显著增强，而非 GPCR 激动剂胶原诱导的血小板聚集和分泌不受影响。我们之前已经证明，GRK6 不参与调控 Gq 偶联的 5HT2A 和 Gz 偶联的 a2A 肾上腺素能受体。有趣的是，与 GRK6 不同的是，5-羟色胺和肾上腺素可分别激活 5-HT2A 和 a2A 肾上腺素能受体，而在 GRK3 -/-血小板中，5-羟色胺和肾上腺素共同刺激诱导的血小板聚集明显增强，这表明 GRK3 参与了一般的 GPCR 调节。此外，在 GRK3 -/-血小板中，血小板对第二次 ADP 挑战的聚集反应得以恢复，而在 WT 血小板中，再次刺激该激动剂未能诱导聚集反应，这证实 GRK3 对一般 GPCR 脱敏做出了贡献。最后，在 GRK3 -/-血小板中，2-MeSADP- 和 AYPGKF 诱导的 AKT 和 ERK 磷酸化显著增强：结论：GRK3 在血小板中通过一般 GPCR 脱敏调节血小板活化过程中起着至关重要的作用。

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来源期刊

Thrombosis and haemostasis 医学-外周血管病

CiteScore

11.90

自引率

9.00%

发文量

140

审稿时长

1 months

期刊介绍： Thrombosis and Haemostasis publishes reports on basic, translational and clinical research dedicated to novel results and highest quality in any area of thrombosis and haemostasis, vascular biology and medicine, inflammation and infection, platelet and leukocyte biology, from genetic, molecular & cellular studies, diagnostic, therapeutic & preventative studies to high-level translational and clinical research. The journal provides position and guideline papers, state-of-the-art papers, expert analysis and commentaries, and dedicated theme issues covering recent developments and key topics in the field.