Highly efficient macromolecule-sized poration of lipid bilayers by a synthetically evolved peptide.
Highly efficient macromolecule-sized poration of lipid bilayers by a synthetically evolved peptide.
复制标题
DOI：
10.1021/ja500462s
复制
发表时间：
2014-03-26
影响因子：
15
通讯作者：
Wimley WC
中科院分区：
文献类型：
--
作者：
Wiedman G;Fuselier T;He J;Searson PC;Hristova K;Wimley WC
在低浓度下自组装成允许大分子通过的跨双层孔的肽将在生物技术的多个领域中是有益的。然而，具有这种性质的天然或设计的肽很少（如果有的话）。在这里，我们表明，26个残基的肽“MelP5”，合成进化的蜂毒裂解肽蜂毒肽的功能获得性变体，确定在高通量筛选小分子泄漏，使大分子通过双层的条件下，蜂毒肽和其他成孔肽不。在表面支撑的双层中，MelP5在低至1：25000的肽：脂质比下形成异常高的电导、平衡孔。由于MelP5引起的双层电导的增加显著高于由于蜂毒肽或其他肽成孔剂的亲本序列引起的增加。在这里，我们还开发了两种新的检测大分子从囊泡泄漏，我们用它们来表征MelP5孔双层。我们表明，MelP5允许通过跨囊泡膜的大分子肽：脂质比低至1：500，并在条件下，既不渗透裂解，也不发生总囊泡不稳定。MelP5形成的大分子大小的平衡孔是独特的，因为在相同条件下蜂毒肽和其他成孔肽都不会显著释放大分子。因此，MelP5似乎属于一种新的功能性肽类，可以形成多种潜在生物技术应用的基础。
Peptides that self-assemble, at low concentration, into bilayer-spanning pores which allow the passage of macromolecules would be beneficial in multiple areas of biotechnology. However, there are few, if any, natural or designed peptides that have this property. Here we show that the 26-residue peptide “MelP5”, a synthetically evolved gain-of-function variant of the bee venom lytic peptide melittin identified in a high-throughput screen for small molecule leakage, enables the passage of macromolecules across bilayers under conditions where melittin and other pore-forming peptides do not. In surface-supported bilayers, MelP5 forms unusually high conductance, equilibrium pores at peptide:lipid ratios as low as 1:25000. The increase in bilayer conductance due to MelP5 is dramatically higher, per peptide, than the increase due to the parent sequence of melittin or other peptide pore formers. Here we also develop two novel assays for macromolecule leakage from vesicles, and we use them to characterize MelP5 pores in bilayers. We show that MelP5 allows the passage of macromolecules across vesicle membranes at peptide:lipid ratios as low as 1:500, and under conditions where neither osmotic lysis nor gross vesicle destabilization occur. The macromolecule-sized, equilibrium pores formed by MelP5 are unique as neither melittin nor other pore-forming peptides release macromolecules significantly under the same conditions. MelP5 thus appears to belong to a novel functional class of peptide that could form the foundation of multiple potential biotechnological applications.
登录
查看更多内容
影响因子：
2.9
作者：
Hristova, K;Selsted, ME;White, SH
通讯作者：
White, SH
影响因子：
64.8
作者：
Cornell, BA;BraachMaksvytis, VLB;Pace, RJ
通讯作者：
Pace, RJ
影响因子：
3.4
作者：
HOPE, MJ;BALLY, MB;CULLIS, PR
通讯作者：
CULLIS, PR
影响因子：
3.4
作者：
DEGRADO, WF;MUSSO, GF;KEZDY, FJ
通讯作者：
KEZDY, FJ
影响因子：
3.4
作者：
Ladokhin, AS;Wimley, WC;White, SH
通讯作者：
White, SH