Evidence that prokineticin receptor 2 exists as a dimer in vivo
Evidence that prokineticin receptor 2 exists as a dimer in vivo
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DOI：
10.1007/s00018-010-0601-6
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发表时间：
2011-09-01
影响因子：
8
通讯作者：
Miele, Rossella
中科院分区：
文献类型：
--
作者：
Marsango, Sara;di Patti, Maria Carmela Bonaccorsi;Miele, Rossella
前动力蛋白是调节多种生物过程的蛋白质，包括胃肠运动、血管生成、昼夜节律和先天免疫应答。前动力素结合两个密切相关的G蛋白偶联受体（GPCR），PKR1和PKR2。一般来说，这些受体作为分子开关将激活传递给异源三聚体G蛋白，越来越多的证据表明GPCR作为同源或异源二聚体存在。我们在这里通过蛋白质印迹分析表明，PKR2在中性粒细胞中具有二聚体结构。通过PKR2在酿酒酵母中的异源表达，我们研究了PKR2二聚化的分子间相互作用机制。三种类型的机制的潜在参与进行了研究：卷曲螺旋，二硫键，和跨膜结构域之间的疏水相互作用。不同的删除或定点PKR2突变体的表征表明，二聚化通过跨膜结构域之间的相互作用进行。我们证明，共表达结合缺陷和信号缺陷形式的PKR2可以重建受体功能，可能是通过结构域交换机制。
Prokineticins are proteins that regulate diverse biological processes including gastrointestinal motility, angiogenesis, circadian rhythm, and innate immune response. Prokineticins bind two closed related G-protein coupled receptors (GPCRs), PKR1 and PKR2. In general, these receptors act as molecular switches to relay activation to heterotrimeric G-proteins and a growing body of evidence points to the fact that GPCRs exist as homo- or heterodimers. We show here by Western-blot analysis that PKR2 has a dimeric structure in neutrophils. By heterologous expression of PKR2 in Saccharomyces cerevisiae, we examined the mechanisms of intermolecular interaction of PKR2 dimerization. The potential involvement of three types of mechanisms was investigated: coiled-coil, disulfide bridges, and hydrophobic interactions between transmembrane domains. Characterization of differently deleted or site-directed PKR2 mutants suggests that dimerization proceeds through interactions between transmembrane domains. We demonstrate that co-expressing binding-deficient and signaling-deficient forms of PKR2 can re-establish receptor functionality, possibly through a domain-swapping mechanism.