KCTD5 Forms Hetero-Oligomeric Complexes with Various Members of the KCTD Protein Family.

KCTD5 Forms Hetero-Oligomeric Complexes with Various Members of the KCTD Protein Family.
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DOI:
10.3390/ijms241814317
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发表时间:
2023-09-20
影响因子:
5.6
通讯作者:
--
中科院分区:
生物学2区
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钾通道四聚化结构域 5 (KCTD5) 调节生理学的各个方面,从神经元信号传导到结直肠癌。 KCTD5 的一个关键特征是它能够自组装成多亚基寡聚物,这似乎能够参与一系列蛋白质-蛋白质相互作用。最近有报道称 KCTD5 与两个相似的 KCTD(KCTD2 和 KCTD17)形成异源寡聚复合物。然而,尚不清楚 KCTD5 是否与包含超过两打成员的其余 KCTD 蛋白家族形成异源寡聚复合物。在这里,我们证明当通过裂解细胞中的免疫共沉淀进行测定时,KCTD5 与各种 KCTD 蛋白相互作用。我们通过检查基于活细胞生物发光共振能量转移 (BRET) 的方法中 KCTD5 的相互作用来强化该数据集。最后,我们开发了一种 IP 发光方法来绘制 KCTD5 上与选定的 KCTD 相互作用所需的区域,这些 KCTD 已在神经元信号传导中发挥作用。我们报告说,KCTD5 上的不同区域负责与其他 KCTD 蛋白的相互作用。虽然我们的结果有助于揭示 KCTD5 的其他相互作用伙伴,但它们也揭示了 KCTD 生物学中的其他复杂性。此外,我们的研究结果还表明,KCTD 异源寡聚相互作用可能发生在整个 KCTD 家族中。
Potassium Channel Tetramerization Domain 5 (KCTD5) regulates diverse aspects of physiology, ranging from neuronal signaling to colorectal cancer. A key feature of KCTD5 is its self-assembly into multi-subunit oligomers that seemingly enables participation in an array of protein–protein interactions. KCTD5 has recently been reported to form hetero-oligomeric complexes with two similar KCTDs (KCTD2 and KCTD17). However, it is not known if KCTD5 forms hetero-oligomeric complexes with the remaining KCTD protein family which contains over two dozen members. Here, we demonstrate that KCTD5 interacts with various KCTD proteins when assayed through co-immunoprecipitation in lysed cells. We reinforced this dataset by examining KCTD5 interactions in a live-cell bioluminescence resonance energy transfer (BRET)-based approach. Finally, we developed an IP-luminescence approach to map regions on KCTD5 required for interaction with a selection of KCTD that have established roles in neuronal signaling. We report that different regions on KCTD5 are responsible for uniquely contributing to interactions with other KCTD proteins. While our results help unravel additional interaction partners for KCTD5, they also reveal additional complexities in KCTDs’ biology. Moreover, our findings also suggest that KCTD hetero-oligomeric interactions may occur throughout the KCTD family.
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