Opitz Robert, Müller Matthias, Reuter Cédric, Barone Matthias, Soicke Arne, Roske Yvette, Piotukh Kirill, Huy Peter, Beerbaum Monika, Wiesner Burkhard, Beyermann Michael, Schmieder Peter, Freund Christian, Volkmer Rudolf, Oschkinat Hartmut, Schmalz Hans-Günther, Kühne Ronald
Leibniz-Institut für Molekulare Pharmakologie, 13125 Berlin, Germany;
Department für Chemie, Universität zu Köln, 50939 Köln, Germany;
Proc Natl Acad Sci U S A. 2015 Apr 21;112(16):5011-6. doi: 10.1073/pnas.1422054112. Epub 2015 Apr 6.
Small-molecule competitors of protein-protein interactions are urgently needed for functional analysis of large-scale genomics and proteomics data. Particularly abundant, yet so far undruggable, targets include domains specialized in recognizing proline-rich segments, including Src-homology 3 (SH3), WW, GYF, and Drosophila enabled (Ena)/vasodilator-stimulated phosphoprotein (VASP) homology 1 (EVH1) domains. Here, we present a modular strategy to obtain an extendable toolkit of chemical fragments (ProMs) designed to replace pairs of conserved prolines in recognition motifs. As proof-of-principle, we developed a small, selective, peptidomimetic inhibitor of Ena/VASP EVH1 domain interactions. Highly invasive MDA MB 231 breast-cancer cells treated with this ligand showed displacement of VASP from focal adhesions, as well as from the front of lamellipodia, and strongly reduced cell invasion. General applicability of our strategy is illustrated by the design of an ErbB4-derived ligand containing two ProM-1 fragments, targeting the yes-associated protein 1 (YAP1)-WW domain with a fivefold higher affinity.
蛋白质-蛋白质相互作用的小分子竞争者对于大规模基因组学和蛋白质组学数据的功能分析至关重要。特别丰富但迄今为止难以成药的靶点包括专门识别富含脯氨酸片段的结构域，如Src同源3（SH3）、WW、GYF以及果蝇Enabled（Ena）/血管舒张刺激磷蛋白（VASP）同源1（EVH1）结构域。在此，我们提出一种模块化策略，以获得一个可扩展的化学片段工具包（ProMs），其设计目的是取代识别基序中保守的脯氨酸对。作为原理验证，我们开发了一种小型、选择性的Ena/VASP EVH1结构域相互作用的拟肽抑制剂。用这种配体处理的高侵袭性MDA MB 231乳腺癌细胞显示，VASP从粘着斑以及片状伪足前端被取代，并且细胞侵袭能力大幅降低。我们通过设计一种含有两个ProM-1片段的ErbB4衍生配体来说明该策略的普遍适用性，该配体以高五倍的亲和力靶向Yes相关蛋白1（YAP1）-WW结构域。
