Identification of progesterone receptor membrane component-1 as an interaction partner and possible regulator of fatty acid 2-hydroxylase
Identification of progesterone receptor membrane component-1 as an interaction partner and possible regulator of fatty acid 2-hydroxylase
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DOI：
10.1042/bcj20170963
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发表时间：
2018-03-15
影响因子：
4.1
通讯作者：
Eckhardt, Matthias
中科院分区：
文献类型：
--
作者：
Hardt, Robert;Winter, Dominic;Eckhardt, Matthias
脂肪酸 2-羟化酶 (FA2H) 对于髓鞘细胞和其他细胞中 2-羟基化脂肪酸的合成至关重要，这种酶的缺乏会导致复杂形式的遗传性痉挛性截瘫，也称为脂肪酸羟化酶相关的神经变性。尽管 FA2H 在鞘脂代谢中发挥重要作用，但对 FA2H 的调节及其与参与相同或其他代谢途径的其他蛋白质的相互作用知之甚少。为了识别酶的潜在相互作用伙伴，使用细胞稳定同位素标记的定量质谱法分别与甲醛交联和邻近生物素化相结合。除了参与鞘脂合成和神经酰胺膜间转移的其他酶以及 FA2H 的假定氧化还原伙伴外，黄体酮受体膜成分 1 (PGRMC1) 和 PGRMC2 也被确定为假定的相互作用伙伴。已知这两种相关的血红素结合蛋白可调节多种细胞色素 P450 酶。双分子荧光互补实验证实了FA2H与PGRMC1的相互作用。此外，PGRMC1 抑制剂 AG-205 显着减少 FA2H 表达细胞中羟基化神经酰胺和葡萄糖神经酰胺的合成。这表明 PGRMC1 可能通过其血红素伴侣活性来调节 FA2H 活性。
The fatty acid 2-hydroxylase (FA2H) is essential for synthesis of 2-hydroxylated fatty acids in myelinating and other cells, and deficiency of this enzyme causes a complicated form of hereditary spastic paraplegia also known as fatty acid hydroxylase-associated neurodegeneration. Despite its important role in sphingolipid metabolism, regulation of FA2H and its interaction with other proteins involved in the same or other metabolic pathways is poorly understood. To identify potential interaction partners of the enzyme, quantitative mass spectrometry using stable isotope labeling of cells was combined with formaldehyde cross-linking and proximity biotinylation, respectively. Besides other enzymes involved in sphingolipid synthesis and intermembrane transfer of ceramide, and putative redox partners of FA2H, progesterone receptor membrane component-1 (PGRMC1) and PGRMC2 were identified as putative interaction partners. These two related heme-binding proteins are known to regulate several cytochrome P450 enzymes. Bimolecular fluorescence complementation experiments confirmed the interaction of FA2H with PGRMC1. Moreover, the PGRMC1 inhibitor AG-205 significantly reduced synthesis of hydroxylated ceramide and glucosylceramide in FA2H-expressing cells. This suggests that PGRMC1 may regulate FA2H activity, possibly through its heme chaperone activity.