<br>pH依赖性β-折叠自组装肽的高通量筛选,Small

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pH依赖性β-折叠自组装肽的高通量筛选
Small ( IF 13.0 ) Pub Date : 2024-01-06 , DOI: 10.1002/smll.202307963
Xin-Wei Ye _{1,

2,

3} , Wen Tian ₁ , Lu Han ₁ , Yi-Jing Li ₁ , Shan Liu ₁ , Wen-Jia Lai ₄ , Yi-Xuan Liu ₁ , Lei Wang ₁ , Pei-Pei Yang ₁ , Hao Wang _{1,

2,

3,

5}

Affiliation

pH 依赖性肽生物材料在细胞输送和组织工程方面具有巨大的潜力。然而，识别具有特定二级结构的响应自组装序列仍然是一个挑战。在这项工作中，开发了一种基于单珠一化合物（OBOC）组合库的实验程序，用于在中性水溶液（pH 7.5）下快速筛选自组装β-折叠肽并在弱酸性条件（pH 6.5）下分解）。使用疏水性荧光分子硫代黄素T（ThT）作为探针，由于ThT插入疏水域，展示自组装肽的树脂珠在pH 7.5下显示荧光，并在pH 6.5溶液中进一步培养。选择荧光熄灭的珠子。鉴定出三种七肽可以在 pH 7.5 下自组装成纳米纤维或纳米颗粒，并在 pH 6.5 下分解。 P1 (LVEFRHY) 通过 pH 调节表现出快速的酸反应和形态转变。组氨酸和苯丙氨酸的疏水性苯基基序的电荷变化可能在pH响应性β-折叠纳米纤维的形成中发挥重要作用。这种高通量筛选方法提供了一种有效的方法来识别 pH 依赖性 β-片层自组装肽，并深入了解此类纳米材料的结构设计。

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High-Throughput Screening of pH-Dependent β-sheet Self-Assembling Peptide

pH-dependent peptide biomaterials hold tremendous potential for cell delivery and tissue engineering. However, identification of responsive self-assembling sequences with specified secondary structure remains a challenge. In this work, An experimental procedure based on the one-bead one-compound (OBOC) combinatorial library is developed to rapidly screen self-assembling β-sheet peptides at neutral aqueous solution (pH 7.5) and disassemble at weak acidic condition (pH 6.5). Using the hydrophobic fluorescent molecule thioflavin T (ThT) as a probe, resin beads displaying self-assembling peptides show fluorescence under pH 7.5 due to the insertion of ThT into the hydrophobic domain, and are further cultured in pH 6.5 solution. The beads with extinguished fluorescence are selected. Three heptapeptides are identified that can self-assemble into nanofibers or nanoparticles at pH 7.5 and disassemble at pH 6.5. P1 (LVEFRHY) shows a rapid acid response and morphology transformation with pH modulation. Changes in the charges of histidine and hydrophobic phenyl motif of phenylalanine may play important roles in the formation of pH-responsive β-sheet nanofiber. This high-throughput screening method provides an efficient way to identify pH-dependent β-sheet self-assembling peptide and gain insights into structural design of such nanomaterials.

更新日期：2024-01-06

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