Thompson David B, Cronican James J, Liu David R
Howard Hughes Medical Institute, Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts, USA.
Methods Enzymol. 2012;503:293-319. doi: 10.1016/B978-0-12-396962-0.00012-4.
Supercharged proteins are a class of engineered or naturally occurring proteins with unusually high positive or negative net theoretical charge. Both supernegatively and superpositively charged proteins exhibit a remarkable ability to withstand thermally or chemically induced aggregation. Superpositively charged proteins are also able to penetrate mammalian cells. Associating cargo with these proteins, such as plasmid DNA, siRNA, or other proteins, can enable the functional delivery of these macromolecules into mammalian cells both in vitro and in vivo. The potency of functional delivery in some cases can exceed that of other current methods for macromolecule delivery, including the use of cell-penetrating peptides such as Tat and adenoviral delivery vectors. This chapter summarizes methods for engineering supercharged proteins, optimizing cell penetration, identifying naturally occurring supercharged proteins, and using these proteins for macromolecule delivery into mammalian cells.
超荷蛋白是一类经过工程改造或天然存在的蛋白质，其理论净电荷具有异常高的正值或负值。带超高负电荷和超高正电荷的蛋白质都表现出显著的抵抗热诱导或化学诱导聚集的能力。带超高正电荷的蛋白质还能够穿透哺乳动物细胞。将货物（如质粒DNA、小干扰RNA或其他蛋白质）与这些蛋白质结合，能够在体外和体内将这些大分子功能性递送至哺乳动物细胞。在某些情况下，功能性递送的效力可能超过目前其他大分子递送方法，包括使用穿膜肽（如Tat）和腺病毒递送载体。本章总结了工程化超荷蛋白的方法、优化细胞穿透的方法、鉴定天然存在的超荷蛋白的方法，以及使用这些蛋白质将大分子递送至哺乳动物细胞的方法。
