A biophysical characterization of the folded domains of KCTD12: insights into interaction with the GABAB2 receptor
A biophysical characterization of the folded domains of KCTD12: insights into interaction with the GABAB2 receptor
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DOI：
10.1002/jmr.2291
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发表时间：
2013-10-01
影响因子：
2.7
通讯作者：
Pedone, Emilia
中科院分区：
文献类型：
--
作者：
Correale, Stefania;Esposito, Carla;Pedone, Emilia
最近的研究表明，KCTD家族的成员在基本的生物过程中发挥重要作用。尽管他们的作用，非常有限的信息是关于他们的结构和分子组织。通过结合不同的实验和理论技术，我们在这里的特点是两个折叠结构域的KCTD 12，一个不可分割的组成部分和GABA（B2）受体的调节剂。二次预测方法和CD光谱表明，N-末端结构域KCTD 12（BTB）假定/结构，而C-末端结构域KCTD 12（H1）的主要特征是由-结构。结合试验表明，两个独立表达的结构域显示出良好的亲和力。这表明整个蛋白质可能具有相当紧凑的结构，具有由长的无序区域连接的两个相互作用的结构域。值得注意的是，KCTD 12（BTB）和KCTD 12（H1）在单独表达时都是四聚体。这一发现可能会改变传统的观点，认为只有POZ/BTB结构域的一个特定的寡聚作用。KCTD 12（POZ/BTB）对GABA（B2）C-末端区域的亲和力的首次定量显示其福尔斯处于低微摩尔范围。有趣的是，我们还证明了GABA（B2）相关肽能够以非常高的亲和力结合KCTD 12（BTB）。该肽可能代表用于调节KCTD 12/GABA（B2）体外相互作用的有用工具，并且也可能构成具有治疗应用潜力的拟肽化合物的开发的起点。版权所有（c）2013约翰威利父子有限公司
Recent investigations have shown that members of the KCTD family play important roles in fundamental biological processes. Despite their roles, very limited information is available on their structures and molecular organization. By combining different experimental and theoretical techniques, we have here characterized the two folded domains of KCTD12, an integral component and modulator of the GABA(B2) receptor. Secondary prediction methods and CD spectroscopy have shown that the N-terminal domain KCTD12(BTB) assumes an / structure, whereas the C-terminal domain KCTD12(H1) is predominantly characterized by a -structure. Binding assays indicate that the two domains independently expressed show a good affinity for each other. This suggests that the overall protein is likely endowed with a rather compact structure with two interacting structured domains joint by a long disordered region. Notably, both KCTD12(BTB) and KCTD12(H1) are tetrameric when individually expressed. This finding could modify the traditional view that ascribes only to POZ/BTB domain a specific oligomerization role. The first quantification of the affinity of KCTD12(POZ/BTB) for the C-terminal region of GABA(B2) shows that it falls in the low micromolar range. Interestingly, we also demonstrate that a GABA(B2)-related peptide is able to bind KCTD12(BTB) with a very high affinity. This peptide may represent a useful tool for modulating KCTD12/GABA(B2) interaction in vitro and may also constitute the starting point for the development of peptidomimetic compounds with a potential for therapeutic applications. Copyright (c) 2013 John Wiley & Sons, Ltd.