Division of Molecular and Cellular Medicine, King Mongkut's Institute of Technology Ladkrabang, Faculty of Medicine, Bangkok, Thailand.
Department of Microbiology, University of Illinois, Urbana, Illinois, United States of America.
PLoS One. 2024 Oct 22;19(10):e0309988. doi: 10.1371/journal.pone.0309988. eCollection 2024.
Aerobic organisms continuously generate internal superoxide and hydrogen peroxide, which can damage enzymes and impair growth. To avoid this problem cells maintain high levels of superoxide dismutases, catalases, and peroxidases. Surprisingly, we do not know the primary sources of these reactive oxygen species (ROS) in living cells. However, in vitro studies have shown that flavoenzymes can inadvertently transfer electrons to oxygen. Therefore, it seems plausible that substantial ROS may be generated when large metabolic fluxes flow through flavoproteins. Such a situation may arise during the catabolism of fatty acids. Acyl-CoA dehydrogenase (FadE) is a flavoprotein involved in each turn of the beta-oxidation cycle. In the present study the catabolism of dodecanoic acid specifically impaired the growth of strains that lack enzymes to scavenge hydrogen peroxide. The defect was absent from fadE mutants. Direct measurements confirmed that the beta-oxidation pathway amplified the rate of intracellular hydrogen peroxide formation. Scavenging-proficient cells did not display the FadE-dependent growth defect. Those cells also did not induce the peroxide stress response during dodecanoate catabolism, indicating that the basal defenses are sufficient to cope with moderately elevated peroxide formation. In vitro work still is needed to test whether the ROS evolve specifically from the FadE flavin site and to determine whether superoxide as well as peroxide is released. At present such experiments are challenging because the natural redox partner of FadE has not been identified. This study supports the hypothesis that the degree of internal ROS production can depend upon the type of active metabolism inside cells.
需氧生物不断产生内部超氧化物和过氧化氢,这会破坏酶并影响生长。为了解决这个问题,细胞维持高水平的超氧化物歧化酶、过氧化氢酶和过氧化物酶。令人惊讶的是,我们不知道这些活性氧物质(ROS)在活细胞中的主要来源。然而,体外研究表明黄素酶可以无意中将电子转移给氧气。因此,当大量代谢通量流经黄素蛋白时,可能会产生大量的 ROS,这似乎是合理的。这种情况可能发生在脂肪酸的分解代谢过程中。酰基辅酶 A 脱氢酶(FadE)是一种参与β-氧化循环每一轮的黄素蛋白。在本研究中,十二烷酸的分解代谢特别损害了缺乏清除过氧化氢酶的菌株的生长。 fadE 突变体没有缺陷。直接测量证实,β-氧化途径放大了细胞内过氧化氢形成的速率。具有清除能力的细胞没有表现出依赖 FadE 的生长缺陷。这些细胞在十二烷酸分解代谢过程中也没有诱导过氧化物应激反应,表明基础防御足以应对适度升高的过氧化物形成。仍需要进行体外工作来测试 ROS 是否是从 FadE 黄素部位特异性产生的,并确定是否释放了超氧化物和过氧化物。目前,由于尚未确定 FadE 的天然氧化还原伴侣,因此此类实验具有挑战性。本研究支持这样一种假设,即内部 ROS 产生的程度可以取决于细胞内活跃代谢的类型。