凋亡对于发育和组织稳态很重要，失调会导致疾病，包括癌症。作为一个凋亡效应物，BAK会发生构象变化促进线粒体外膜破裂，导致细胞死亡，这被称为“活化”，可以由人类蛋白BID，BIM和PUMA产生的肽诱发。为了鉴定能够调节BAK的额外肽，我们使用了计算蛋白设计，酵母表面显示筛选和基于结构能量评分来鉴定10种不同的新结合物。我们发现了来自人类蛋白BNIP5和PXT1以及三种非天然肽，它们在脂质体实验中激活BAK并引起细胞色素C从线粒体释放。晶体结构和结合研究揭示了肽激动剂和抑制剂之间的高度相似性，排除了简单的功能决定性属性。我们的研究结果揭示了可以调控BAK功能的巨大肽序列空间，并将指导BAK调节工具和治疗药物的设计。版权所有 © 2023 Elsevier Ltd. 保留所有权利。

Apoptosis is important for development and tissue homeostasis, and its dysregulation can lead to diseases, including cancer. As an apoptotic effector, BAK undergoes conformational changes that promote mitochondrial outer membrane disruption, leading to cell death. This is termed "activation" and can be induced by peptides from the human proteins BID, BIM, and PUMA. To identify additional peptides that can regulate BAK, we used computational protein design, yeast surface display screening, and structure-based energy scoring to identify 10 diverse new binders. We discovered peptides from the human proteins BNIP5 and PXT1 and three non-native peptides that activate BAK in liposome assays and induce cytochrome c release from mitochondria. Crystal structures and binding studies reveal a high degree of similarity among peptide activators and inhibitors, ruling out a simple function-determining property. Our results shed light on the vast peptide sequence space that can regulate BAK function and will guide the design of BAK-modulating tools and therapeutics.Copyright © 2023 Elsevier Ltd. All rights reserved.