被磷酸化激活的混合谱系激酶域样蛋白(MLKL)被发现转移到质膜，MLKL通过位于其n端区域的四螺旋束结构域(4HBD)表面的带正电荷氨基酸片与磷脂酰肌醇磷酸盐(PIPs)相互作用(Dondelinger Y et al. 2014;Wang H et al. 2014;Hildebrand JM et al. 2014;Su L et al. 2014;Quarato G et al. 2016)。使用PIKfyve (P5i)等PIP结合物干扰PI(5)P或PI(4,5)P2的形成，可以有效地抑制小鼠L929和人fadd缺失Jurkat细胞中tnf诱导的坏死(Dondelinger Y et al. 2014)。体外脂质体实验显示，MLKL可诱导含PIP或心磷脂脂质体的泄漏，这表明MLKL可能具有成孔能力，通过细胞膜的通透性介导细胞死亡(Dondelinger Y et al. 2014;Wang H et al. 2014;Tanzer MC et al. 2016; Petrie EJ et al. 2018). Liposome permeabilization assays demonstrated that the N-terminal 4HB domain of MLKL compromised membrane integrity, and was more effective on liposomes whose composition resembled that of plasma membranes than on those mimicking mitochondrial membranes (Tanzer MC et al. 2016). One study has proposed the 4HB domain might reorganize in membranes to assemble into ion channels (Xia B et al. 2016); however, this remains to be fully explored in cellular contexts and structurally. Other studies implicated MLKL in engaging mitochondrial membranes to provoke mitochondrial fission or promote ion channel activity (Cai Z et al. 2014), although subsequent studies have discounted these possibilities (Murphy JM et al. 2013; Tait SW et al. 2013; Moujalled DM et al. 2014; Wang H et al. 2014; reviewd by Murphy JM 2020). Studies in human cell lines suggest that upon induction of necroptosis MLKL shifts to the plasma membrane and membranous organelles such as mitochondria, lysosome, endosome and ER (Wang H et al. 2014), but MLKL trafficking via a Golgi-microtubule-actin-dependent mechanism facilitates plasma membrane accumulation of MLKL, where membrane disruption causes death (Samson AL et al. 2020). Based on studies showing that the 4HB domain can permeabilize membranes in vitro (Dondelinger Y et al. 2014; Su L et al. 2014; Wang H et al. 2014; Tanzer MC et al. 2016; Petrie EJ et al. 2018), it is thought that MLKL kills cells via direct action on the plasma membrane (Murphy JM 2020).

多项研究表明，运输途径所需的内体分选复合物(ESCRT)可以从坏死细胞中去除磷酸化的含mlkl的膜泡，从而减弱细胞死亡过程(Gong YN et al. 2017;Yoon S et al. 2017;Zargarian S et al. 2017;Fan W et al. 2019)。escrt相关蛋白，程序性细胞死亡6相互作用蛋白(PDCD6IP或alg -2相互作用蛋白X, ALIX)和syntenin-1 (SDCBP)被发现可拮抗mlkl介导的质膜改变(Fan W et al. 2019)。此外，有人提出了flotillin介导的内吞作用，通过从质膜上去除MLKL并将其重新导向溶酶体降解来抑制坏死下垂(Fan W et al. 2019)。