Prosaposin regulates coenzyme Q10 levels in HepG2 cells, especially those in mitochondria.
Prosaposin regulates coenzyme Q10 levels in HepG2 cells, especially those in mitochondria.
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DOI：
10.3164/jcbn.13-106
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发表时间：
2014-09
影响因子：
2.4
通讯作者：
Yamamoto Y
中科院分区：
文献类型：
--
作者：
Kashiba M;Oizumi M;Suzuki M;Sawamura Y;Nagashima K;Yoshimura S;Yamamoto Y
辅酶Q10 （CoQ10）是线粒体电子传递链的关键组成部分，是最重要的细胞抗氧化剂之一。我们以前报道过糖蛋白皂苷B （SapB）在人类细胞中与辅酶q10结合。为了阐明SapB及其前体prosaposin （Psap）的生理作用，我们制备了稳定的HepG2过表达野生型人Psap （Wt-Tf）的转染物。我们还建立了一个SapB结构域突变的Psap (Mt-Tf)，其中半胱氨酸198被丝氨酸取代，通过失去S-S桥接来破坏三维蛋白质结构。Psap敲低（KD）菌株也进行了检测。Western blotting分析证实Psap在HepG2细胞中过表达或低表达。CoQ10与游离胆固醇（FC）的细胞比值从Wt-Tf>亲本>Mt-Tf>KD的顺序显著降低。此外，线粒体组分中CoQ10/FC的比值按照Wt-Tf>亲本>KD的顺序依次降低。这些数据表明Psap和/或SapB调节HepG2细胞中的CoQ10水平，特别是在其线粒体中。
Coenzyme Q10 (CoQ10) is a key component of the mitochondrial electron transfer chain and is one of the most important cellular antioxidants. We previously reported that glycoprotein saposin B (SapB) binds CoQ10 in human cells. To elucidate the physiological role of SapB and its precursor, prosaposin (Psap), we prepared stable transfectants of HepG2 that overexpress wild-type human Psap (Wt-Tf). We also established a SapB domain mutated Psap (Mt-Tf) in which cysteine198 was replaced with serine to disrupt three dimensional protein structure by the loss of S-S bridging. Psap knockdown (KD) strains were also examined. Western blotting analysis confirmed overexpression or knockdown of Psap in these HepG2 cells. The cellular ratios of CoQ10 to free cholesterol (FC) significantly decreased in the order of Wt-Tf>parental>Mt-Tf>KD. Additionally, the ratios of CoQ10/FC in mitochondrial fractions decreased in the order of Wt-Tf>parental>KD. These data indicate that Psap and/or SapB regulate CoQ10 levels in HepG2 cells, especially in their mitochondria.
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影响因子：
2.4
作者：
Jin, GuangZhi;Horinouchi, Ryo;Yamamoto, Yorihiro
通讯作者：
Yamamoto, Yorihiro
影响因子：
1.9
作者：
KALEN, A;APPELKVIST, EL;DALLNER, G
通讯作者：
DALLNER, G
影响因子：
2.8
作者：
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通讯作者：
Vessella, Robert L.
影响因子：
8
作者：
Ahn, Victoria E.;Leyko, Paul;Prive, Gilbert G.
通讯作者：
Prive, Gilbert G.
影响因子：
3.9
作者：
ABERG, F;APPELKVIST, EL;ERNSTER, L
通讯作者：
ERNSTER, L