Paralog-selective ligands for Bcl-2 proteins
Paralog-selective ligands for Bcl-2 proteins
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DOI：
10.1021/ja0441211
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发表时间：
2005-02-16
影响因子：
15
通讯作者：
Schepartz, A
中科院分区：
文献类型：
--
作者：
Gemperli, AC;Rutledge, SE;Schepartz, A
目前，结合细胞内或细胞外蛋白质表面并抑制蛋白质-蛋白质相互作用的分子具有相当大的兴趣，以前我们已经报道了基于胰腺折叠多肽的微型蛋白质可以识别即使是浅的α-螺旋结合裂缝，对不相关的蛋白质具有高亲和力和选择性。PPBH 3 -1是一种这样的微型蛋白，它以纳摩尔亲和力与抗凋亡蛋白旁系同源物Bcl-2和Bcl-XL结合，并且对Bcl-XL的亲和力为ΔΔG= 1.2 kcal·mol-1。在这里，我们描述了PPBH 3 -1的定向进化成两个新的微型蛋白质，PPBH 3 -5和PPBH 3 -6，其parabolic特异性相对于PPBH 3 -1是反向的。PPBH 3 -5和PPBH 3 -6以纳摩尔亲和力结合Bcl-2，并且相对于Bcl-XL，对Bcl-2的ΔΔG= 0.9−1.3 kcal·mol-1偏好。用Bcl-XL变体进行的实验表明，PPBH 3 -5和PPBH 3 -6通过利用Bcl-2和Bcl-XL分子景观中的细微结构或静电差异来实现高的特异性。PPBH 3 -5和PPBH 3 -6可能作为与Bcl-2蛋白选择性相互作用的非天然配体的早期例子具有独特的应用。
There is considerable current interest in molecules that bind intra- or extracellular protein surfaces and inhibit protein−protein interactions.Previously we have reported that miniature proteins based on pancreatic-fold polypeptides can recognize even shallow α-helix binding clefts with high affinity and selectivity against unrelated proteins. One such miniature protein, PPBH3-1, binds the anti-apoptotic protein paralogs Bcl-2 and Bcl-XLwith nanomolar affinity and a ΔΔG= 1.2 kcal·mol-1preference for Bcl-XL. Here we describe the directed evolution of PPBH3-1 into two new miniature proteins, PPBH3-5 and PPBH3-6, whose paralog specificity is reversed relative to PPBH3-1. PPBH3-5 and PPBH3-6 bind Bcl-2 with nanomolar affinity and a ΔΔG= 0.9−1.3 kcal·mol-1preference for Bcl-2 over Bcl-XL. Experiments with Bcl-XLvariants suggest that PPBH3-5 and PPBH3-6 achieve high paralog specificity by exploiting subtle structural or electrostatic differences in the Bcl-2 and Bcl-XLmolecular landscapes. PPBH3-5 and PPBH3-6 may have unique applications as early examples of nonnatural ligands that interact selectively with Bcl-2 proteins.