Institute of Molecular Biosciences, Mahidol University, 25/25 Phuttamonthon 4 Road, Salaya, Phuttamonthon, Nakhon Pathom 73170, Thailand
Institute of Molecular Biosciences, Mahidol University, 25/25 Phuttamonthon 4 Road, Salaya, Phuttamonthon, Nakhon Pathom 73170, Thailand.
Biol Open. 2020 Apr 28;9(4):bio051110. doi: 10.1242/bio.051110.
Previously, we have developed an extramitochondrial assembly system, where mitochondrial targeting signal (MTS) can be removed from a given mitochondrial enzyme, which could be used to characterize the regulatory factors involved in enzyme assembly/disassembly Here, we demonstrate that addition of exogenous acetaldehyde can quickly induce the supramolecular assembly of MTS-deleted aldehyde dehydrogenase Ald4p in yeast cytoplasm. Also, by using PCR-based modification of the yeast genome, cytoplasmically targeted Ald4p cannot polymerize into long filaments when key functional amino acid residues are substituted, as shown by N192D, S269A, E290K and C324A mutations. This study has confirmed that extramitochondrial assembly could be a powerful external system for studying mitochondrial enzyme assembly, and its regulatory factors outside the mitochondria. In addition, we propose that mitochondrial enzyme assembly/disassembly is coupled to the regulation of a given mitochondrial enzyme activity.
先前,我们开发了一种线粒体外组装系统,在此系统中,线粒体靶向信号(MTS)可从给定的线粒体酶中去除,这可用于鉴定参与酶组装/拆卸的调节因子。在这里,我们证明了外加乙醛可快速诱导 MTS 缺失的醛脱氢酶 Ald4p 在酵母细胞质中的超分子组装。此外,通过基于 PCR 的酵母基因组修饰,当关键功能氨基酸残基被取代时,靶向细胞质的 Ald4p 不能聚合成长纤维,如 N192D、S269A、E290K 和 C324A 突变所示。这项研究证实了线粒体外组装可以成为研究线粒体酶组装及其在线粒体外调节因子的强大外部系统。此外,我们提出线粒体酶的组装/拆卸与特定线粒体酶活性的调节相偶联。