TAT-RasGAP_317–326 is a cell-penetrating peptide-based construct with anticancer and antimicrobial activities. This peptide kills a subset of cancer cells in a manner that does not involve known programmed cell death pathways. Here we have elucidated the mode of action allowing TAT-RasGAP_317–326 to kill cells. This peptide binds and disrupts artificial membranes containing lipids typically enriched in the inner leaflet of the plasma membrane, such as phosphatidylinositol-bisphosphate (PIP₂) and phosphatidylserine (PS). Decreasing the amounts of PIP₂ in cells renders them more resistant to TAT-RasGAP_317–326, while reducing the ability of cells to repair their plasma membrane makes them more sensitive to the peptide. The W317A TAT-RasGAP_317–326 point mutant, known to have impaired killing activities, has reduced abilities to bind and permeabilize PIP₂- and PS-containing membranes and to translocate through biomembranes, presumably because of a higher propensity to adopt an α-helical state. This work shows that TAT-RasGAP_317–326 kills cells via a form of necrosis that relies on the physical disruption of the plasma membrane once the peptide targets specific phospholipids found on the cytosolic side of the plasma membrane.

TAT-rasGAP的_317-326是具有抗癌和抗微生物活性的细胞穿透肽基构建体。该肽以不涉及已知的程序性细胞死亡途径的方式杀死癌细胞的子集。在这里，我们已经阐明的行动让TAT-rasGAP的模式_317-326杀死细胞。该肽结合并破坏人造膜，所述人造膜包含通常富含质膜内部小叶的脂质，例如磷脂酰肌醇双磷酸酯（PIP ₂）和磷脂酰丝氨酸（PS）。降低PIP的量₂在细胞使得它们到TAT-rasGAP的更耐_317-326，同时降低细胞修复其质膜的能力，使其对肽更敏感。所述W317A TAT-rasGAP的_317-326位点突变体，已知有受损的杀伤活性，降低的能力结合和透PIP ₂ -和含PS膜和易位通过生物膜，大概是因为更高的倾向采用的α-螺旋状态。这项工作表明，TAT-rasGAP的_317-326经由坏死的一种形式，依赖于质膜一旦肽靶的特异性磷脂在质膜的胞质侧发现的物理破坏杀死细胞。