Protein cell-surface display through in situ enzymatic modification of proteins with a poly(Ethylene glycol)-lipid.
Protein cell-surface display through in situ enzymatic modification of proteins with a poly(Ethylene glycol)-lipid.
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通过用聚乙二醇脂质对蛋白质进行原位酶促修饰来展示蛋白质细胞表面。
DOI：
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发表时间：
2013
影响因子：
3.8
通讯作者：
Teruyuki Nagamune
中科院分区：
文献类型：
--
作者：
U. Tomita;S. Yamaguchi;Yasukazu Maeda;Kazuki Chujo;K. Minamihata;Teruyuki Nagamune
功能蛋白质的细胞表面展示是调节和增强细胞功能的有力和有用的工具。直接掺入来自细胞外介质的位点特异性脂化蛋白质比通过基因转移表达在展示活性蛋白质方面更快速、更容易控制和更可靠。然而，需要不期望的两亲性试剂如有机共溶剂和去污剂来抑制溶液中普通脂化蛋白质的聚集。在这里，我们报告分选酶A催化的蛋白质修饰与聚乙二醇（PEG）-脂质在活细胞表面上原位。与识别标签融合的蛋白质与初步掺入细胞膜的PEG-脂质位点特异性连接。因此，在无两亲性试剂的条件下，靶蛋白成功地展示在活细胞上而没有聚集。此外，为了证明本方法的可用性，在癌细胞上展示免疫球蛋白G的Fc结构域，并且通过Fc-Fc受体相互作用增强癌细胞与树突细胞的吞噬作用。因此，本发明的用于蛋白质展示的简便的化学酶促方法可用于调节细胞和组织工程领域中的细胞-细胞相互作用。
Cell-surface display of functional proteins is a powerful and useful tool for regulating and reinforcing cellular functions. Direct incorporation of site-specifically lipidated proteins from the extracellular medium is more rapid, easily controllable and reliable in displaying active proteins than expression through gene transfer. However, undesirable amphiphilic reagents such as organic co-solvents and detergents were required for suppressing aggregation of ordinary lipidated proteins in solution. We report here sortase A-catalyzed modification of proteins with a poly(ethylene glycol)(PEG)-lipid in situ on the surface of living cells. Proteins fused with a recognition tag were site-specifically ligated with the PEG-lipid which was preliminary incorporated into cell membranes. Accordingly, target proteins were successfully displayed on living cells without aggregation under an amphiphilic reagent-free condition. Furthermore, to demonstrate the availability of the present method, Fc domains of immunoglobulin G were displayed on cancer cells, and the phagocytosis of cancer cells with dendritic cells were enhanced through the Fc-Fc receptor interaction. Thus, the present facile chemoenzymatic method for protein display can be utilized for modulating cell-cell interactions in cell and tissue engineering fields.
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使用分子酶A介导的转换的蛋白质对脂质体进行特定位点特异性修饰的新方法。
影响因子：
4.7
作者：
Guo, Xueqing;Wu, Zhimeng;Guo, Zhongwu
通讯作者：
Guo, Zhongwu
影响因子：
15
作者：
Rabuka, David;Forstner, Martin B.;Bertozzi, Carolyn R.
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Bertozzi, Carolyn R.
通过分类酶催化的转肽对蛋白质的脂质修饰。
影响因子：
15
作者：
Antos, John M.;Miller, Gwenn M.;Grotenbreg, Gijsbert M.;Ploegh, Hidde L.
通讯作者：
Ploegh, Hidde L.
影响因子：
56.9
作者：
Saxon, E;Bertozzi, CR
通讯作者：
Bertozzi, CR