Affinity-based screening of peptides recognizing assembly states of self-assembling peptide nanomaterials.
Affinity-based screening of peptides recognizing assembly states of self-assembling peptide nanomaterials.
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DOI：
10.1021/ja905250u
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发表时间：
2009-09
影响因子：
15
通讯作者：
Toshiki Sawada;Tsuyoshi Takahashi;H. Mihara
中科院分区：
文献类型：
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作者：
Toshiki Sawada;Tsuyoshi Takahashi;H. Mihara
从随机七肽库中筛选出能够与自组装肽纳米材料结合的新型肽。针对由β-折叠肽构建的螺旋卷曲纳米纤维的亲和力依赖性肽筛选给出了展示具有各种亲和力和选择性的肽的噬菌体克隆。使用噬菌体展示肽的酶联免疫吸附测定显示，筛选的肽特异性地结合到目标纳米纤维，与由具有略微不同的氨基酸序列的肽组成的参考纳米纤维相反。使用化学合成的肽的斑点印迹测定证明，具有序列Thr-Gly-Val-Lys-Gly-Pro-Gly的肽01（p01）显示出对靶纳米纤维的亲和常数（3.7 × 10（5）M（-1）），其亲和常数比其对单体肽的亲和常数大200倍，比其对短纳米纤维的亲和常数大60倍。这些结果表明，p01选择性地识别靶肽的组装状态。ATR/IR二级结构分析清楚地表明，当p01与靶纳米纤维结合时，它经历了从无规卷曲到平行β-折叠结构的结构转变，导致对靶纤维的更大亲和力和高特异性。通过p01对肽纳米纤维的表面改性证明肽特异性结合到纳米纤维的边缘。使用p01，可以在肽纳米材料上产生金纳米颗粒（蛋白质）的均匀阵列。
Novel peptides capable of binding to self-assembled peptide nanomaterials were identified from a random heptapeptide library displayed on phages. Affinity-dependent peptide screening against helically coiled nanofibers constructed of beta-sheet peptides gave phage clones displaying peptides with a variety of affinities and selectivities. An enzyme-linked immunosorbent assay using phage-displayed peptides revealed that the screened peptides specifically bind to target nanofibers, in contrast to reference nanofibers comprised of peptides with slightly different amino acid sequences. A Dot blot assay using chemically synthesized peptides demonstrated that peptide 01 (p01), with the sequence Thr-Gly-Val-Lys-Gly-Pro-Gly, showed an affinity constant (3.7 x 10(5) M(-1)) for the target nanofibers 200 times greater than its affinity for monomeric peptides and 60 times greater than for short nanofibers. These results suggested that p01 selectively recognizes the assembly state of the target peptide. ATR/IR secondary structure analyses clearly showed that when p01 binds to target nanofibers, it undergoes a structural transition from random-coil to parallel beta-sheet structures, resulting in greater affinity and high specificity for the target fiber. Surface modification of the peptide nanofibers by p01 demonstrated that the peptide specifically binds to the edge of the nanofibers. Using p01, uniform arrays of gold nanoparticles (proteins) could be generated on the peptide nanomaterials.