Duke University, Department of Neurology, USA.
Duke University, Department of Neurology, USA.
Biochim Biophys Acta Mol Basis Dis. 2017 Nov;1863(11):2973-2986. doi: 10.1016/j.bbadis.2017.07.031. Epub 2017 Jul 30.
A variable-length poly-T variant in intron 6 of the TOMM40 gene, rs10524523, is associated with risk and age-of-onset of sporadic (late-onset) Alzheimer's disease. In Caucasians, the three predominant alleles at this locus are Short (S), Long (L) or Very long (VL). On an APOE ε3/3 background, the S/VL and VL/VL genotypes are more protective than S/S. The '523 poly-T has regulatory properties, in that the VL poly-T results in higher expression than the S poly-T in luciferase expression systems. The aim of the current work was to identify effects on cellular bioenergetics of increased TOM40 protein expression. MitoTracker Green fluorescence and autophagic vesicle staining was the same in control and over-expressing cells, but TOM40 over-expression was associated with increased expression of TOM20, a preprotein receptor of the TOM complex, the mitochondrial chaperone HSPA9, and PDHE1a, and increased activities of the oxidative phosphorylation complexes I and IV and of the TCA member α-ketoglutaric acid dehydrogenase. Consistent with the complex I findings, respiration was more sensitive to inhibition by rotenone in control cells than in the TOM40 over-expressing cells. In the absence of inhibitors, total cellular ATP, the mitochondrial membrane potential, and respiration were elevated in the over-expressing cells. Spare respiratory capacity was greater in the TOM40 over-expressing cells than in the controls. TOM40 over-expression blocked Ab-elicited decreases in the mitochondrial membrane potential, cellular ATP levels, and cellular viability in the control cells. These data suggest elevated expression of TOM40 may be protective of mitochondrial function.
载脂蛋白 E ε3/3 背景下,S/VL 和 VL/VL 基因型比 S/S 基因型更具保护作用。“523 多态 T 具有调节特性,即 VL 多态 T 导致荧光素酶表达系统中 S 多态 T 的表达更高。目前这项工作的目的是确定增加 TOM40 蛋白表达对细胞生物能量学的影响。在对照和过表达细胞中,MitoTracker Green 荧光和自噬囊泡染色相同,但 TOM40 过表达与 TOM20 表达增加有关,TOM20 是 TOM 复合物的前蛋白受体、线粒体伴侣 HSPA9 和 PDHE1a,以及氧化磷酸化复合物 I 和 IV 和 TCA 成员α-酮戊二酸脱氢酶的活性增加。与复合物 I 的发现一致,与过表达 TOM40 的细胞相比,对照细胞中呼吸对鱼藤酮的抑制更敏感。在没有抑制剂的情况下,过表达细胞中的总细胞 ATP、线粒体膜电位和呼吸作用升高。过表达 TOM40 的细胞的备用呼吸能力大于对照细胞。TOM40 过表达阻止了 Ab 诱导的对照细胞中线粒体膜电位、细胞 ATP 水平和细胞活力的降低。这些数据表明,TOM40 的高表达可能对线粒体功能具有保护作用。