AlphaFold version 2.0 elucidates the binding mechanism between VIPR2 and KS-133, and reveals an S-S bond (Cys25-Cys192) formation of functional significance for VIPR2
AlphaFold version 2.0 elucidates the binding mechanism between VIPR2 and KS-133, and reveals an S-S bond (Cys25-Cys192) formation of functional significance for VIPR2
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DOI：
10.1016/j.bbrc.2022.10.071
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发表时间：
2022-10-28
影响因子：
3.1
通讯作者：
Hirokawa, Takatsugu
中科院分区：
文献类型：
--
作者：
Sakamoto, Kotaro;Asano, Satoshi;Hirokawa, Takatsugu
血管活性肠肽受体2 （VIPR2）作为治疗精神障碍、癌症和免疫疾病的药物靶点引起了人们的关注。在2021年，我们确定了肽KS-133作为vipr2选择性拮抗剂。在本研究中，我们旨在阐明VIPR2与KS-133的结合机制。为此，通过AlphaFold 2.0版本和分子动力学模拟，构建VIPR2/KS-133和VIPR2/血管活性肠肽（vasactive intestinal peptide， VIP）复合物模型。我们的模型显示：(i) KS-133和VIP都具有螺旋结构，（ii）两种肽在VIPR2上的相互作用残基相似，（iii）它们与VIPR2结合时的螺旋方向相差约45度。有趣的是，在构建上述模型的过程中，人类VIPR2的Cys（25）和Cys（192）之间形成了S-S键。虽然这两种Cys残基在物种间高度保守（即对应于小鼠中的Cys（24）和Cys(191)），但此前没有关于这种S-S键形成的报道。为了阐明这一S-S键在VIPR2中潜在的作用是否具有功能后果，我们生成了表达小鼠VIPR2的细胞系（Cys24Ala, Cys191Ala）。在VIP刺激该细胞系的过程中，发现VIPR2的下游信号标志物AKT的磷酸化明显减弱，从而提示S-S键对VIPR2具有功能意义。我们的研究不仅首次阐明了KS-133的VIPR2结合机制，而且为VIPR2的结构生物学研究提供了新的见解。(c) 2022爱思唯尔公司版权所有。
The vasoactive intestinal peptide receptor 2 (VIPR2) has attracted attention as a drug target for the treatment of mental disorders, cancer, and immune diseases. In 2021, we identified the peptide KS-133 as a VIPR2-selective antagonist. In this study, we aimed to elucidate the binding mechanism between VIPR2 and KS-133. To this end, VIPR2/KS-133 and VIPR2/vasoactive intestinal peptide (VIP) complex models were constructed through AlphaFold version 2.0 and molecular dynamic simulations. Our models revealed that: (i) both KS-133 and VIP have helical structures, (ii) the interaction residues on VIPR2 for both peptides are similar, and (iii) the orientation of their helices upon their binding to VIPR2 are different by similar to 45 degrees. Interestingly, in the process of constructing the aforementioned models, an S-S bond formation between Cys(25) and Cys(192) of the human VIPR2 was identified. Although these two Cys residues are highly conserved among species (i.e., corresponding to Cys(24) and Cys(191) in the mouse), no previous reports regarding this S-S bond formation exist. In order to clarify the potential role of this S-S bond in the VIPR2 has functional consequences, a cell line expressing the mouse VIPR2(Cys24Ala, Cys191Ala) was generated. During the VIP stimulation of this cell line, the phosphorylation of AKT (a downstream signal marker of VIPR2) was found to be significantly attenuated, thereby suggesting that the S-S bond has a functional significance for VIPR2. Our study not only elucidates the VIPR2-binding mechanism of KS-133 for the first time, but also provides new insights into the structural biology of VIPR2. (c) 2022 Elsevier Inc. All rights reserved.