Design of Lipid?Protein Conjugates Using Amphiphilic Peptide Substrates of Microbial Transglutaminase
Design of Lipid?Protein Conjugates Using Amphiphilic Peptide Substrates of Microbial Transglutaminase
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使用微生物转谷氨酰胺酶的两亲性肽底物设计脂质？蛋白质缀合物
DOI：
10.1021/acsabm.8b00271
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发表时间：
2018
影响因子：
4.7
通讯作者：
Kamiya Noriho
中科院分区：
文献类型：
--
作者：
Takahara Mari;Wakabayashi Rie;Minamihata Kosuke;Goto Masahiro;Kamiya Noriho
蛋白质的脂质修饰在调节细胞环境中起着重要作用。模拟天然脂化蛋白质是评估脂质修饰蛋白质功能的关键技术，也是使脂质自组装成靶蛋白的关键技术。在此，我们报告了一种简便的方法，共轭蛋白质与脂质融合肽在均匀的生理条件下，通过使用微生物转氨酶（MTG）反应。MTG催化蛋白质中的特定谷氨酰胺（Q）与新设计的脂质融合肽中的赖氨酸（K）之间的交联反应。新合成了用于脂质修饰的水溶性肽底物C14-X-MRHKGS，其中C14、X和MRHKGS分别代表肉豆蔻酸、由G、P或S组成的接头肽以及被碱性氨基酸包围的MTG反应性K。MTG介导的在C-末端与LLQG融合的蛋白质和溶解在磷酸盐溶液中的C14-X-MRHKGS（5摩尔当量）之间的交联反应产生产率为70至100%的脂质-蛋白质缀合物。所得脂质-蛋白质缀合物对细胞膜的锚定能力依赖于GnS接头中G残基的数目，表明GnS基序的自组装和疏水性用于增强脂质-蛋白质缀合物的膜锚定。
Lipid modification of proteins plays a significant role in regulating the cellular environment. Mimicking natural lipidated proteins is a key technique for assessing the function of proteins modified with lipids and also to render self-assembly of lipids to a target protein. Herein, we report a facile method of conjugating proteins with lipid-fused peptides under homogeneous physiological conditions by using the microbial transglutaminase (MTG) reaction. MTG catalyzes the cross-linking reaction between a specific glutamine (Q) in a protein and a lysine (K) in newly designed lipid-fused peptides. The water-soluble peptide substrates for lipid modification, C14-X-MRHKGS, were newly synthesized, where C14, X, and MRHKGS represent myristic acid, linker peptides composed of G, P, or S, and MTG-reactive K surrounded with basic amino acids, respectively. The MTG-mediated cross-linking reaction between a protein fused with LLQG at the C-terminus and C14-X-MRHKGS (5 molar eq) dissolved in a phosphate saline solution resulted in lipid–protein conjugates with yields of 70 to 100%. The anchoring ability of the obtained lipid–protein conjugates to cell membranes was dependent on the number of G residues in the GnS linker, suggesting that self-assembly and hydrophobicity of the GnS motif serves to enhance membrane anchoring of lipid–protein conjugates.