Structural and thermodynamical insights into the binding and inhibition of FIH-1 by the N-terminal disordered region of Mint3.
Structural and thermodynamical insights into the binding and inhibition of FIH-1 by the N-terminal disordered region of Mint3.
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DOI：
10.1016/j.jbc.2021.101304
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发表时间：
2021-11
期刊：
影响因子：
--
通讯作者：
Tsumoto K
中科院分区：
文献类型：
--
作者：
Ten T;Nagatoishi S;Maeda R;Hoshino M;Nakayama Y;Seiki M;Sakamoto T;Tsumoto K
已知Mint 3通过与FIH-1结合来增强有氧ATP产生，称为瓦尔堡效应。由于这种作用被认为对癌细胞有益，因此这种相互作用是癌症治疗的一个有希望的靶点。然而，先前的研究表明，Mint 3与FIH-1的相互作用区域本质上是无序的，这使得研究这种相互作用具有挑战性。因此，我们采用溶液中的热力学和结构研究来阐明Mint 3与FIH-1结合的结构和化学变化。首先，使用圆二色谱，核磁共振和氢/氘交换质谱（HDX-MS）的组合，我们证实了N-末端的一半，这是Mint 3的相互作用部分，主要是无序的。接下来，我们揭示了一个大的焓和熵变化的相互作用Mint 3使用等温滴定量热法（ITC）。该曲线与无序Mint 3的柔性在与FIH-1结合后急剧降低的模型一致。此外，我们用几种截短的Mint 3进行了一系列ITC实验，这是一种有效的方法，因为Mint 3的相互作用部分是无序的，并将氨基酸78至88鉴定为与FIH-1结合的新核心位点。Mint 3的截断研究还揭示了Mint 3的每个部分对与FIH-1相互作用的热力学贡献，其中核心位点通过形成非共价键对亲和力（ΔG）做出贡献，而其他位点仅影响焓（ΔH）。这一见解可以作为进一步研究内在无序区（IDR）和癌症治疗药物开发的立足点。
Mint3 is known to enhance aerobic ATP production, known as the Warburg effect, by binding to FIH-1. Since this effect is considered to be beneficial for cancer cells, the interaction is a promising target for cancer therapy. However, previous research has suggested that the interacting region of Mint3 with FIH-1 is intrinsically disordered, which makes investigation of this interaction challenging. Therefore, we adopted thermodynamic and structural studies in solution to clarify the structural and thermodynamical changes of Mint3 binding to FIH-1. First, using a combination of circular dichroism, nuclear magnetic resonance, and hydrogen/deuterium exchange–mass spectrometry (HDX-MS), we confirmed that the N-terminal half, which is the interacting part of Mint3, is mostly disordered. Next, we revealed a large enthalpy and entropy change in the interaction of Mint3 using isothermal titration calorimetry (ITC). The profile is consistent with the model that the flexibility of disordered Mint3 is drastically reduced upon binding to FIH-1. Moreover, we performed a series of ITC experiments with several types of truncated Mint3s, an effective approach since the interacting part of Mint3 is disordered, and identified amino acids 78 to 88 as a novel core site for binding to FIH-1. The truncation study of Mint3 also revealed the thermodynamic contribution of each part of Mint3 to the interaction with FIH-1, where the core sites contribute to the affinity (ΔG), while other sites only affect enthalpy (ΔH), by forming noncovalent bonds. This insight can serve as a foothold for further investigation of intrinsically disordered regions (IDRs) and drug development for cancer therapy.
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影响因子：
4.6
作者：
Nakaoka HJ;Hara T;Yoshino S;Kanamori A;Matsui Y;Shimamura T;Sato H;Murakami Y;Seiki M;Sakamoto T
通讯作者：
Sakamoto T
影响因子：
3.9
作者：
Potter M;Newport E;Morten KJ
通讯作者：
Morten KJ
影响因子：
5.7
作者：
Sakamoto T;Seiki M
通讯作者：
Seiki M
DOI：
10.1073/pnas.0509217103
发表时间：
2006-01-31
影响因子：
11.1
作者：
Dietz, H;Rief, M
通讯作者：
Rief, M
影响因子：
64.5
作者：
Radhakrishnan, I;PerezAlvarado, GC;Wright, PE
通讯作者：
Wright, PE