State Key Laboratory of Urban Water Resource and Environment, School of Chemistry and Chemical Engineering, Harbin Institute of Technology, 92 West Da-Zhi Street, Harbin 150001, China.
College of Materials and Chemical Engineering, Heilongjiang Institute of Technology, 999 Hongqi Street, Harbin 150050, China.
Anal Chem. 2022 Mar 8;94(9):3811-3818. doi: 10.1021/acs.analchem.1c04696. Epub 2022 Feb 21.
Mass transport across cell membranes is a primary process for cellular metabolism. For this purpose, electrostatically mediated membrane fusion is exploited to transport various small molecules including glucose-6-phosphate, isopropyl β-D-thiogalactoside, and macromolecules such as DNA plasmids from negatively charged large unilamellar vesicles (LUVs) to positively charged giant unilamellar vesicles (GUVs). After membrane fusion between these oppositely charged vesicles, molecules are transported into GUVs to trigger the NAD involved enzyme reaction, bacterial gene expression, and in vitro gene expression of green fluorescent protein from a DNA plasmid. The optimized charged lipid percentages are 10% for both positively charged GUVs and negatively charged LUVs to ensure the fusion process. The experimental results demonstrate a universal way for mass transport into the artificial cells through vesicle fusions, which paves a crucial step for the investigation of complicated cellular metabolism.
跨细胞膜的物质运输是细胞代谢的主要过程。为此，利用静电介导的膜融合将各种小分子，包括葡萄糖-6-磷酸、异丙基β-D-硫代半乳糖苷以及大分子如 DNA 质粒，从带负电荷的大单室脂质体 (LUVs) 运输到带正电荷的巨单室脂质体 (GUVs)。在这些带相反电荷的囊泡之间发生膜融合后，分子被运入 GUVs 以触发 NAD 相关的酶反应、细菌基因表达以及从 DNA 质粒体外表达绿色荧光蛋白。优化的带电荷脂质百分比为正电荷 GUVs 和负电荷 LUVs 各 10%，以确保融合过程的进行。实验结果证明了通过囊泡融合将物质大规模输送到人工细胞中的通用方法，为研究复杂的细胞代谢铺平了关键的一步。
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