Extending the Half-Life of a Protein in Vivo by Enzymatic Labeling with Amphiphilic Lipopeptides
Extending the Half-Life of a Protein in Vivo by Enzymatic Labeling with Amphiphilic Lipopeptides
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通过两亲性脂肽酶标记延长体内蛋白质的半衰期
DOI：
10.1021/acs.bioconjchem.1c00027
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发表时间：
2021
影响因子：
4.7
通讯作者：
Noriho Kamiya
中科院分区：
文献类型：
--
作者：
Mari Takahara ; Shinichi Mochizuki ; Rie Wakabayashi ; Kosuke Minamihata ; Masahiro Goto ; Kazuo Sakurai ; Noriho Kamiya
脂蛋白偶联物的合成是蛋白质药物传递系统中的重要技术之一，然而由于脂类分子的疏水性，脂蛋白的完整偶联仍然具有挑战性。为了便于处理结合中的脂质部分，我们重点研究了一种微生物谷氨酰胺转氨酶(MTG)，它可以连接脂肽和感兴趣蛋白质中的特定赖氨酸(K)和谷氨酰胺(Q)残基。作为MTG底物，设计了单脂和双脂融合的两亲性短脂多肽底物(Lid-G3S-RHK或lipid2-KG3S-RHK)。这些两亲性脂肽和一种与LLQG融合的模型蛋白(增强型绿色荧光蛋白)(LQ-EGFP)都是水溶性的，因此即使使用胆固醇-G3S-RHK，通过MTG反应也可以有效地获得LQ-EGFP转化率为80%的脂-蛋白结合物。体外细胞黏附和体内半衰期稳定性的评价表明，单胆固醇-G3S-RHK修饰的蛋白质通过形成稳定的白蛋白/脂-蛋白复合体而获得最高的细胞黏附效率和最长的半衰期。
Synthesis of lipid–protein conjugates is one of the significant techniques in drug delivery systems of proteins; however, the intact conjugation of a lipid and protein is yet challenging due to the hydrophobicity of lipid molecules. In order to facilitate easy handling of the lipid moiety in conjugation, we have focused on a microbial transglutaminase (MTG) that can ligate specific lysine (K) and glutamine (Q) residues in lipopeptides and a protein of interest. As MTG substrates, monolipid- and dilipid-fused amphiphilic short lipopeptide substrates (lipid-G3S-RHK or lipid2-KG3S-RHK) were designed. These amphiphilic lipopeptides and a model protein (enhanced green fluorescent protein, EGFP) fused with LLQG (LQ-EGFP) were both water-soluble, and thus lipid–protein conjugates were efficiently obtained through the MTG reaction with a >80% conversion rate of LQ-EGFP even using cholesterol-G3S-RHK.In vitrocell adhesion andin vivohalf-life stability of the successfully obtained lipid–protein conjugates were evaluated, showing that the monocholesterol-G3S-RHK modification of a protein gave the highest cell adhesion efficiency and longest half-life time by formation of a stable albumin/lipid–protein complex.