Structural model for the interaction of a designed Ankyrin Repeat Protein with the human epidermal growth factor receptor 2.
Structural model for the interaction of a designed Ankyrin Repeat Protein with the human epidermal growth factor receptor 2.
复制标题
DOI：
10.1371/journal.pone.0059163
复制
发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Adams TE
中科院分区：
文献类型：
--
作者：
Epa VC;Dolezal O;Doughty L;Xiao X;Jost C;Plückthun A;Adams TE
设计的锚蛋白重复序列蛋白是一类新型结合蛋白，其可以被选择和进化以高亲和力和特异性结合靶标。我们对DARPin H10-2-G3感兴趣，该DARPin H10-2-G3已经进化为以非常高的亲和力与人表皮生长因子受体2（HER 2）结合。研究发现HER 2在30%的乳腺癌中过度表达，并且是FDA批准的治疗性单克隆抗体曲妥珠单抗和帕妥珠单抗以及小分子酪氨酸激酶抑制剂的靶点。在这里，我们使用计算大分子对接，加上几个接口度量，如形状互补性，相互作用能，和静电互补性，模型的DARPin H10-2-G3和HER 2之间的复合物的结构。我们分析了两种蛋白质之间的界面，然后通过显示在与H10-2-G3的假定界面处选择的HER 2点突变使结合亲和力降低高达100倍而不影响曲妥珠单抗的结合来验证结构模型。与随后解决的X-射线晶体结构的配合物的比较得到的骨架原子的均方根偏差为0.84-1.14毫微克。这里提出的研究表明，结构生物信息学的计算技术在生成有用的蛋白质-蛋白质相互作用的结构模型的能力。
Designed Ankyrin Repeat Proteins are a class of novel binding proteins that can be selected and evolved to bind to targets with high affinity and specificity. We are interested in the DARPin H10-2-G3, which has been evolved to bind with very high affinity to the human epidermal growth factor receptor 2 (HER2). HER2 is found to be over-expressed in 30% of breast cancers, and is the target for the FDA-approved therapeutic monoclonal antibodies trastuzumab and pertuzumab and small molecule tyrosine kinase inhibitors. Here, we use computational macromolecular docking, coupled with several interface metrics such as shape complementarity, interaction energy, and electrostatic complementarity, to model the structure of the complex between the DARPin H10-2-G3 and HER2. We analyzed the interface between the two proteins and then validated the structural model by showing that selected HER2 point mutations at the putative interface with H10-2-G3 reduce the affinity of binding up to 100-fold without affecting the binding of trastuzumab. Comparisons made with a subsequently solved X-ray crystal structure of the complex yielded a backbone atom root mean square deviation of 0.84–1.14 Ångstroms. The study presented here demonstrates the capability of the computational techniques of structural bioinformatics in generating useful structural models of protein-protein interactions.
登录
查看更多内容
影响因子：
14.9
作者：
Schymkowitz J;Borg J;Stricher F;Nys R;Rousseau F;Serrano L
通讯作者：
Serrano L
DOI：
10.1073/pnas.89.8.3338
发表时间：
1992-04-15
影响因子：
11.1
作者：
NORTHRUP, SH;ERICKSON, HP
通讯作者：
ERICKSON, HP
影响因子：
5.6
作者：
Binz, HK;Stumpp, MT;Plückthun, A
通讯作者：
Plückthun, A
影响因子：
16
作者：
Garrett, TPJ;McKern, NM;Ward, CW
通讯作者：
Ward, CW
影响因子：
50.3
作者：
Junttila, Teemu T.;Akita, Robert W.;Sliwkowski, Mark X.
通讯作者：
Sliwkowski, Mark X.