Biomolecular engineering by combinatorial design and high-throughput screening: small, soluble peptides that permeabilize membranes.
Biomolecular engineering by combinatorial design and high-throughput screening: small, soluble peptides that permeabilize membranes.
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DOI：
10.1021/ja8017863
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发表时间：
2008-07-30
影响因子：
15
通讯作者：
Wimley, William C.
中科院分区：
文献类型：
--
作者：
Rathinakumar, Ramesh;Wimley, William C.
膜活性肽的合理设计和工程化在很大程度上仍然是一个未被满足的目标。我们假设，这部分是由于某些膜活性，如渗透作用，不依赖于特定的氨基酸序列或特定的三维肽结构。相反，它们依赖于界面活动；分子进入膜-水界面并改变脂质堆积和组织的能力。在这里，我们通过采用非经典方法进行生物分子工程和膜活性肽的设计来测试这一想法。从全长9-15个氨基酸的16,384个理性组合肽库中筛选出可渗透磷脂膜的可溶性成员。采用严格的两相高通量筛选，鉴定出10种具有强膜渗透活性的独特肽，但也是水溶性的。这些罕见而独特的活性肽不具有任何特定的序列基序、肽长或净电荷，而是具有共同的组成特征、二级结构和核心疏水性。我们表明，它们的作用主要取决于界面活性的共同机制，并导致瞬态孔隙形成。我们在这里证明了组合空间肽库与基于功能的高通量筛选相结合可以导致发现多样化，可溶性和高效的膜渗透肽。
Rational design and engineering of membrane active peptides remains a largely unsatisfied goal. We have hypothesized that this us due, in part, to the fact that some membrane activities, such as permeabilization, are not dependent on specific amino acid sequences or specific three-dimensional peptide structures. Instead they depend on interfacial activity; the ability of a molecule to partition into in the membrane-water interface and to alter the packing and organization of lipids. Here we test that idea by taking a non-classical approach to biomolecular engineering and design of membrane-active peptides. A 16,384 member rational combinatorial peptide library, containing peptides 9-15 amino acids in length, was screened for soluble members that permeabilize phospholipid membranes. A stringent, two-phase, high-throughput screen was used to identify 10 unique peptides that had potent membrane permeabilizing activity, but were also water soluble. These rare and uniquely active peptides did not share any particular sequence motif, peptide length or net charge, but instead share common compositional features, secondary structure and core hydrophobicity. We show that they function by a common mechanism that depends mostly on interfacial activity, and leads to transient pore formation. We demonstrate here that composition-space peptide libraries coupled with function-based high-throughput screens can lead to the discovery of diverse, soluble, and highly potent membrane permeabilizing peptides.
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