Specific binding of GM1-binding peptides to high-density GM1 in lipid membranes
Specific binding of GM1-binding peptides to high-density GM1 in lipid membranes
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DOI：
10.1021/la0619067
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发表时间：
2007-01-16
期刊：
影响因子：
3.9
通讯作者：
Sato, Toshinori
中科院分区：
文献类型：
--
作者：
Matsubara, Teruhiko;Iijima, Kazutoshi;Sato, Toshinori
神经节苷脂Gal β 1-3GalNAc β 1-4（Neu 5Ac α 2-3）Gal β 1-4Glc β 1- 1 'Cer（GM 1）是一种重要的受体。我们以前确定了GM 1结合肽的基础上亲和选择从随机肽库。在本研究中，我们确定了结合GM 1所必需的氨基酸，并研究了与GM 1在脂质膜中的特异性相互作用。共有序列（W/F）RxL（xP/Px）xFxx（Rx/xR）xP中的精氨酸和芳香族氨基酸有助于肽结合GM 1的能力。具有共有序列的肽p3，VWRLLAPPFSNRLLP，显示出对GM 1的高亲和力，解离常数为1.2 μ M。此外，使用GM 1和Glc β 1- 1 'Cer（GlcCer）的混合单层研究了p3的密度依赖性结合。p3优先结合到高密度的GM 1，和它的相互作用与GM 1被发现是合作的基础上的希尔图。这些结果表明，GM 1分子的横向组装是由p3识别碳水化合物所必需的。GM 1结合肽作为不同于霍乱毒素B亚基或抗体的独特抗GM 1探针发挥作用。
The ganglioside Gal beta 1-3GalNAc beta 1-4(Neu5Ac alpha 2-3)Gal beta 1-4Glc beta 1-1'Cer (GM1) is an important receptor. We have previously identified GM1-binding peptides based on affinity selection from a random peptide library. In the present study, we determined the amino acids essential for binding GM1 and investigated the specific interaction with GM1 in the lipid membrane. Arginines and aromatic amino acids in the consensus sequence (W/F)RxL(xP/Px)xFxx(Rx/xR)xP contributed to the ability of the peptides to bind GM1. The peptide p3, VWRLLAPPFSNRLLP, having the consensus sequence, showed high affinity for GM1 with a dissociation constant of 1.2 mu M. Furthermore, the density-dependent binding of p3 was investigated using mixed monolayers of GM1 and Glc beta 1-1'Cer (GlcCer). p3 binds preferentially to high-density GM1, and its interaction with GM1 was found to be cooperative based on a Hill plot. These results indicated that a lateral assembly of GM1 molecules was required for the recognition of carbohydrates by p3. The GM1-binding peptide played a role as a unique anti-GM1 probe differing from the cholera toxin B subunit or antibodies.