Nonnatural protein-protein interaction-pair design by key residues grafting
文献类型：期刊论文
| 作者 | Sen Liu; Liu, Shiyong; Zhu, Xiaolei; Liang, Huanhuan; Cao, Aoneng; Chang, Zhijie; Lai, Luhua |
| 刊名 | PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
 |
| 出版日期 | 2007-03-27 |
| 卷号 | 104期号:13页码:5330-5335 |
| 关键词 | De Novo Design Of Protein-protein Interaction Pair Erythropoietin Functional Site Grafting Key Residue At Interface |
| ISSN号 | 0027-8424 |
| DOI | 10.1073/pnas.0606198104 |
| 英文摘要 | Protein-protein interface design is one of the most exciting fields in protein science; however, designing nonnatural protein-protein interaction pairs remains difficult. In this article we report a de novo design of a nonnatural protein-protein interaction pair by scanning the Protein Data Bank for suitable scaffold proteins that can be used for grafting key interaction residues and can form stable complexes with the target protein after additional mutations. Using our design algorithm, an unrelated protein, rat PLC delta(1)-PH (pleckstrin homology domain of phospholipase C-delta 1), was successfully designed to bind the human erythropoietin receptor (EPOR) after grafting the key interaction residues of human erythropoietin binding to EPOR. The designed mutants of rat PLC delta(1)-PH were expressed and purified to test their binding affinities with EPOR. A designed triple mutation of PLC delta(1)-PH (ERPH1) was found to bind EPOR with high affinity (K(D) of 24 nM and an IC(50) of 5.7 mu M) both in vitro and in a cell-based assay, respectively, although the WT PLC delta(1)-PH did not show any detectable binding under the assay conditions. The in vitro binding affinities of the PLC delta(1)-PH mutants correlate qualitatively to the computational binding affinities, validating the design and the protein-protein interaction model. The successful practice of finding a proper protein scaffold and making it bind with EPOR demonstrates a prospective application in protein engineering targeting protein-protein interfaces. |
| 语种 | 英语 |
| WOS记录号 | WOS:000245331700016 |
| 出版者 | NATL ACAD SCIENCES |
| 源URL | [http://ir.iccas.ac.cn/handle/121111/60927]  |
| 专题 | 中国科学院化学研究所 |
| 通讯作者 | Lai, Luhua |
| 作者单位 | 1.Peking Univ, Beijing Natl Lab Mol Sci, State Key Lab Struct Chem Unstable & Stable Speci, Coll Chem & Mol Engn, Beijing 100871, Peoples R China 2.Peking Univ, Ctr Theoret Biol, Beijing 100871, Peoples R China 3.Tsinghua Univ, Sch Med, Dept Biol Sci & Biotechnol, Beijing 100084, Peoples R China |
| 推荐引用方式 GB/T 7714 | Sen Liu,Liu, Shiyong,Zhu, Xiaolei,et al. Nonnatural protein-protein interaction-pair design by key residues grafting[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,2007,104(13):5330-5335. |
| APA | Sen Liu.,Liu, Shiyong.,Zhu, Xiaolei.,Liang, Huanhuan.,Cao, Aoneng.,...&Lai, Luhua.(2007).Nonnatural protein-protein interaction-pair design by key residues grafting.PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,104(13),5330-5335. |
| MLA | Sen Liu,et al."Nonnatural protein-protein interaction-pair design by key residues grafting".PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 104.13(2007):5330-5335. |
入库方式： OAI收割
来源：化学研究所
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