George Nathalie, Pick Horst, Vogel Horst, Johnsson Nils, Johnsson Kai
Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédérale de Lausanne (EPFL), CH-1015 Lausanne, Switzerland.
J Am Chem Soc. 2004 Jul 28;126(29):8896-7. doi: 10.1021/ja048396s.
The specific and covalent labeling of fusion proteins with synthetic molecules opens up new ways to study protein function in the living cell. Here we present a novel method that allows for the specific and exclusive extracellular labeling of proteins on the surfaces of live cells with a large variety of synthetic molecules including fluorophores, protein ligands, or quantum dots. The approach is based on the specific labeling of fusion proteins of acyl carrier protein with synthetic molecules through post-translational modification catalyzed by phosphopantetheine transferase. The specificity and versatility of the labeling should allow it to become an important tool for studying and manipulating cell surface proteins and for complementing existing approaches in cell surface engineering.
用合成分子对融合蛋白进行特异性共价标记为研究活细胞中的蛋白质功能开辟了新途径。在此,我们提出了一种新方法,该方法能够用包括荧光团、蛋白质配体或量子点在内的多种合成分子对活细胞表面的蛋白质进行特异性和排他性的细胞外标记。该方法基于通过磷酸泛酰巯基乙胺转移酶催化的翻译后修饰,用合成分子对酰基载体蛋白的融合蛋白进行特异性标记。标记的特异性和多功能性应使其成为研究和操纵细胞表面蛋白以及补充细胞表面工程中现有方法的重要工具。