Labeling cell surface glycosylphosphatidylinositol-anchored proteins through metabolic engineering using an azide-modified phosphatidylinositol.
Labeling cell surface glycosylphosphatidylinositol-anchored proteins through metabolic engineering using an azide-modified phosphatidylinositol.
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DOI：
10.1016/j.bbrc.2023.01.029
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发表时间：
2023-02-19
影响因子：
3.1
通讯作者：
Guo, Zhongwu
中科院分区：
文献类型：
--
作者：
Kundu, Sayan;Jaiswal, Mohit;Craig, Kendall C.;Guo, Jiatong;Guo, Zhongwu
关键词：
糖基磷脂酰肌醇（GPI）锚定是真核细胞质膜上最常见的蛋白附着机制之一。gpi锚定蛋白（GPI-APs）在许多生物过程中起着至关重要的作用，但很难研究。本研究利用叠氮修饰的磷脂酰肌醇（PI）作为细胞表面gpi - ap的生物合成前体，建立了一种有效的、选择性的代谢工程和标记方法。结果表明，该叠氮化物- pi衍生物被HeLa细胞吸收并结合到gpi的生物合成途径中，在活细胞表面呈现叠氮化物标记的GPI-APs。叠氮基团作为分子手柄，通过生物相容性点击反应安装其他标签，实现荧光成像、蛋白下拉等多种生物学研究，有助于探索gpi - ap的功能，发现新的gpi - ap。
Glycosylphosphatidylinositol (GPI) anchorage is one of the most common mechanisms to attach proteins to the plasma membrane of eukaryotic cells. GPI-anchored proteins (GPI-APs) play a critical role in many biological processes but are difficult to study. Here, a new method was developed for the effective and selective metabolic engineering and labeling of cell surface GPI-APs with an azide-modified phosphatidylinositol (PI) as the biosynthetic precursor of GPIs. It was demonstrated that this azido-PI derivative was taken up by HeLa cells and incorporated into the biosynthetic pathway of GPIs to present azide-labeled GPI-APs on the live cell surface. The azido group was used as a molecular handle to install other labels through a biocompatible click reaction to enable various biological studies, e.g., fluorescent imaging and protein pull-down, which can help explore the functions of GPI-APs and discover new GPI-APs.
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影响因子：
56.9
作者：
FERGUSON, MAJ;HOMANS, SW;RADEMACHER, TW
通讯作者：
RADEMACHER, TW
影响因子：
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作者：
Mehlert, Angela;Ferguson, Michael A. J.
通讯作者：
Ferguson, Michael A. J.
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Varón Silva D
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影响因子：
4.7
作者：
Hussein NH;Amin NS;El Tayebi HM
通讯作者：
El Tayebi HM
影响因子：
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作者：
Elortza, F;Nühse, TS;Jensen, ON
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