Defunctionalizing intracellular organelles such as mitochondria and peroxisomes with engineered phospholipase A/acyltransferases.
Defunctionalizing intracellular organelles such as mitochondria and peroxisomes with engineered phospholipase A/acyltransferases.
复制标题
DOI：
10.1038/s41467-022-31946-5
复制
发表时间：
2022-07-29
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
作者：
细胞器具有维持细胞稳态的多方面功能。操纵单个细胞器的遗传学和药理学方法在探索它们的生理作用方面是强大的。然而，它们中的许多要么行动缓慢，仅限于某些细胞器，要么依赖于有毒物质。在这里，我们设计了一个通用的分子工具，利用磷脂酶A/酰基转移酶（PLAAT）的细胞器通过重塑膜磷脂的快速去功能化。特别是，我们确定催化活性PLAAT截断最小的不利特性。优化的PLAAT向线粒体表面的化学诱导易位导致其以磷脂酶活性依赖性方式快速变形，随后是管腔蛋白的损失以及膜电位的耗散，从而使功能失效。为了证明广泛的适用性，我们然后在过氧化物酶体中调整分子工具，并观察基质驻留功能蛋白的泄漏。该技术与原代神经元培养物中的光遗传学控制、病毒递送和操作相容。由于这种多功能性，PLAAT策略应该证明是有用的，在研究不同背景下的细胞器生物学。操纵单个细胞器的方法对于了解它们的功能非常重要。在这里，作者报告了一种利用磷脂酶A/酰基转移酶（PLAAT）通过重塑膜磷脂快速去功能化细胞器的工具。
Organelles vitally achieve multifaceted functions to maintain cellular homeostasis. Genetic and pharmacological approaches to manipulate individual organelles are powerful in probing their physiological roles. However, many of them are either slow in action, limited to certain organelles, or rely on toxic agents. Here, we design a generalizable molecular tool utilizing phospholipase A/acyltransferases (PLAATs) for rapid defunctionalization of organelles via remodeling of the membrane phospholipids. In particular, we identify catalytically active PLAAT truncates with minimal unfavorable characteristics. Chemically-induced translocation of the optimized PLAAT to the mitochondria surface results in their rapid deformation in a phospholipase activity dependent manner, followed by loss of luminal proteins as well as dissipated membrane potential, thus invalidating the functionality. To demonstrate wide applicability, we then adapt the molecular tool in peroxisomes, and observe leakage of matrix-resident functional proteins. The technique is compatible with optogenetic control, viral delivery and operation in primary neuronal cultures. Due to such versatility, the PLAAT strategy should prove useful in studying organelle biology of diverse contexts. Approaches for manipulating individual organelles are important for learning more about their functions. Here the authors report a tool utilising phospholipase A/acyltransferases (PLAATs) for rapid defunctionalisation of organelles through remodelling of the membrane phospholipids.
登录
查看更多内容
影响因子：
21.3
作者：
Bleazard, W;McCaffery, JM;Shaw, JM
通讯作者：
Shaw, JM
影响因子：
3.4
作者：
Greiner, Jack, V;Glonek, Thomas
通讯作者：
Glonek, Thomas
膜结合细胞器的形态和功能。
影响因子：
7.5
作者：
Heald, Rebecca;Cohen-Fix, Orna
通讯作者：
Cohen-Fix, Orna
影响因子：
7.7
作者：
Itoh, Kie;Murata, Daisuke;Sesak, Hiromi
通讯作者：
Sesak, Hiromi
影响因子：
48
作者：
Bindels, Daphne S.;Haarbosch, Lindsay;Gadella, Theodorus W. J., Jr.
通讯作者：
Gadella, Theodorus W. J., Jr.