Verteporfin is a substrate-selective γ-secretase inhibitor that binds the amyloid precursor protein transmembrane domain.
Verteporfin is a substrate-selective γ-secretase inhibitor that binds the amyloid precursor protein transmembrane domain.
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DOI：
10.1016/j.jbc.2022.101792
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发表时间：
2022-04
期刊：
影响因子：
--
通讯作者：
Sanders CR
中科院分区：
文献类型：
--
作者：
Castro MA;Parson KF;Beg I;Wilkinson MC;Nurmakova K;Levesque I;Voehler MW;Wolfe MS;Ruotolo BT;Sanders CR
这项工作报告了基于小分子抑制剂与底物的直接结合，对具有广泛底物特异性的蛋白酶进行底物选择性抑制。这些研究的目标是γ-分泌酶蛋白酶，它能裂解数十种不同的单跨膜蛋白底物，包括人类淀粉样蛋白前体蛋白的C99结构域和Notch受体。需要对 C99 裂解进行底物特异性抑制，以减少淀粉样蛋白-β 多肽的产生，而不抑制 Notch 裂解，Notch 裂解是与广泛特异性 γ-分泌酶抑制剂相关的主要毒性来源。为了鉴定人类 γ-分泌酶的 C99 选择性抑制剂，我们对模型膜中 FDA 批准的针对 C99 的药物进行了基于 NMR 的筛选。从这个屏幕中，我们鉴定出了具有这些特性的小分子维替泊芬。我们观察到维替泊芬与 C99 形成直接 1:1 复合物，KD 为 15–47 μM（取决于所使用的膜模拟物），并且它不结合 Notch-1 受体的跨膜结构域。生化测定表明，维替泊芬与 C99 的直接结合可抑制 C99 的 γ-分泌酶裂解，IC50 值在 15–164 μM 范围内，而 Notch-1 裂解仅在较高浓度下受到抑制，并且可能是通过不涉及与 Notch-1 结合的机制来抑制。这项工作记录了一种基于 NMR 的稳健方法，用于发现单跨膜蛋白的小分子结合物，并证实可以以底物特异性方式抑制 γ-分泌酶。
This work reports substrate-selective inhibition of a protease with broad substrate specificity based on direct binding of a small-molecule inhibitor to the substrate. The target for these studies was γ-secretase protease, which cleaves dozens of different single-span membrane protein substrates, including both the C99 domain of the human amyloid precursor protein and the Notch receptor. Substrate-specific inhibition of C99 cleavage is desirable to reduce production of the amyloid-β polypeptide without inhibiting Notch cleavage, a major source of toxicity associated with broad specificity γ-secretase inhibitors. In order to identify a C99-selective inhibitors of the human γ-secretase, we conducted an NMR-based screen of FDA-approved drugs against C99 in model membranes. From this screen, we identified the small-molecule verteporfin with these properties. We observed that verteporfin formed a direct 1:1 complex with C99, with a KD of 15–47 μM (depending on the membrane mimetic used), and that it did not bind the transmembrane domain of the Notch-1 receptor. Biochemical assays showed that direct binding of verteporfin to C99 inhibits γ-secretase cleavage of C99 with IC50 values in the range of 15–164 μM, while Notch-1 cleavage was inhibited only at higher concentrations, and likely via a mechanism that does not involve binding to Notch-1. This work documents a robust NMR-based approach to discovery of small-molecule binders to single-span membrane proteins and confirmed that it is possible to inhibit γ-secretase in a substrate-specific manner.
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影响因子：
4.3
作者：
Alford RF;Koehler Leman J;Weitzner BD;Duran AM;Tilley DC;Elazar A;Gray JJ
通讯作者：
Gray JJ
影响因子：
5.3
作者：
Bulic B;Ness J;Hahn S;Rennhack A;Jumpertz T;Weggen S
通讯作者：
Weggen S
影响因子：
4.8
作者：
Donohue, Elizabeth;Tovey, Andrew;Roberge, Michel
通讯作者：
Roberge, Michel
影响因子：
4.2
作者：
Bourgeois, Alexandre;Lauritzen, Inger;Pardossi-Piquard, Raphaelle
通讯作者：
Pardossi-Piquard, Raphaelle
影响因子：
2.9
作者：
Deatherage, Catherine L.;Lu, Zhenwei;Sanders, Charles R.
通讯作者：
Sanders, Charles R.