Department of Chemical and Biological University of Colorado, Boulder, USA.
Chem Commun (Camb). 2020 May 2;56(35):4820-4823. doi: 10.1039/c9cc09401f. Epub 2020 Apr 1.
Efficient intracellular delivery of biomacromolecules such as proteins continues to remain a challenge despite its potential for medicine. In this work, we show that mScarlet, a non cytotoxic red fluorescent protein (RFP) conjugated to Click Nucleic Acid (CNA), a synthetic analog of DNA, undergo cell uptake significantly more than either native proteins or proteins conjugated with similar amounts of DNA in MDA-MB-468 cells. We further demonstrate that the process of cell uptake is metabolically driven and that scavenger receptors and caveolae mediated endocytosis play a significant role. Co-localization studies using anti-scavenger receptor antibodies suggest that scavenger receptors are implicated in the mechanism of uptake of CNA modified proteins.
尽管具有医学应用潜力，但生物大分子（如蛋白质）的高效细胞内递依然是一个挑战。在这项工作中，我们表明，与天然蛋白或与等量 DNA 偶联的蛋白相比，与点击核酸（CNA）偶联的非细胞毒性红色荧光蛋白（RFP）mScarlet 在 MDA-MB-468 细胞中的摄取量显著增加。我们进一步证明，细胞摄取的过程是代谢驱动的，并且清道夫受体和 caveolae 介导的内吞作用发挥了重要作用。使用抗清道夫受体抗体的共定位研究表明，清道夫受体参与了 CNA 修饰蛋白的摄取机制。
