Exploratory Metabolomics Underscores the Folate Enzyme ALDH1L1 as a Regulator of Glycine and Methylation Reactions.
Exploratory Metabolomics Underscores the Folate Enzyme ALDH1L1 as a Regulator of Glycine and Methylation Reactions.
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DOI：
10.3390/molecules27238394
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发表时间：
2022-12-01
期刊：
影响因子：
--
通讯作者：
Krupenko SA
中科院分区：
文献类型：
--
作者：
Rushing BR;Fogle HM;Sharma J;You M;McCormac JP;Molina S;Sumner S;Krupenko NI;Krupenko SA
叶酸（维生素B 9）参与单碳转移反应，在核酸合成和细胞增殖控制等关键细胞过程中发挥重要作用。现在人们认识到，叶酸在癌症发生的不同阶段中的作用是复杂的，需要更多的研究来了解叶酸反应如何在癌症中失调以及由此发生的代谢后果。ALDH 1 L1（胞质10-甲酰四氢叶酸脱氢酶）是一种在许多组织中表达的叶酸代谢酶，在癌症中普遍下调，在癌细胞系中不表达。RT 4细胞系（来自乳头状膀胱癌）表达高水平的ALDH 1 L1代表一个例外，提供了一个机会，探索这种酶的损失的代谢后果。我们已经在RT 4细胞中下调了这种蛋白质（shRNA驱动的敲除或CRISPR驱动的敲除），并比较了ALDH 1 L1表达细胞和ALDH 1 L1缺陷细胞的代谢组，以确定与这种酶缺失相关的代谢变化是否可能为癌细胞提供增殖和/或存活优势。在本研究中，使用超高效液相色谱高分辨率质谱法（UHPLC-HR-MS）分析细胞提取物。使用内部图书馆和公共数据库共识别或注释了13，339个信号。监督和无监督的多变量分析显示RT 4细胞和ALDH 1 L1缺陷克隆之间的代谢差异。甘氨酸（8倍减少）和代谢产物来源于S-腺苷甲硫氨酸利用途径显着减少ALDH 1 L1缺陷克隆，与RT 4细胞相比。与ALDH 1 L1下调相关的其他变化包括氨基酸、克雷布斯循环中间体和核糖-5-磷酸水平降低以及烟酸水平升高。虽然ALDH 1 L1催化的反应与甘氨酸生物合成和甲基通量直接相关，但其对细胞代谢的总体影响超出了由该酶控制的直接代谢途径。
Folate (vitamin B9) is involved in one-carbon transfer reactions and plays a significant role in nucleic acid synthesis and control of cellular proliferation, among other key cellular processes. It is now recognized that the role of folates in different stages of carcinogenesis is complex, and more research is needed to understand how folate reactions become dysregulated in cancers and the metabolic consequences that occur as a result. ALDH1L1 (cytosolic 10-formyltetrahydrofolate dehydrogenase), an enzyme of folate metabolism expressed in many tissues, is ubiquitously downregulated in cancers and is not expressed in cancer cell lines. The RT4 cell line (derived from papillary bladder cancer) which expresses high levels of ALDH1L1 represents an exception, providing an opportunity to explore the metabolic consequences of the loss of this enzyme. We have downregulated this protein in RT4 cells (shRNA driven knockdown or CRISPR driven knockout) and compared metabolomes of ALDH1L1-expressing and -deficient cells to determine if metabolic changes linked to the loss of this enzyme might provide proliferative and/or survival advantages for cancer cells. In this study, cell extracts were analyzed using Ultra High Performance Liquid Chromatography High Resolution Mass Spectrometry (UHPLC-HR-MS). A total of 13,339 signals were identified or annotated using an in-house library and public databases. Supervised and unsupervised multivariate analysis revealed metabolic differences between RT4 cells and ALDH1L1-deficient clones. Glycine (8-fold decrease) and metabolites derived from S-adenosylmethionine utilizing pathways were significantly decreased in the ALDH1L1-deficient clones, compared with RT4 cells. Other changes linked to ALDH1L1 downregulation include decreased levels of amino acids, Krebs cycle intermediates, and ribose-5-phosphate, and increased nicotinic acid. While the ALDH1L1-catalyzed reaction is directly linked to glycine biosynthesis and methyl group flux, its overall effect on cellular metabolism extends beyond immediate metabolic pathways controlled by this enzyme.
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影响因子：
46.9
作者：
通讯作者：
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影响因子：
5
作者：
Hu, Zhenrong;Yang, Ranran;Hu, Jingzhou
通讯作者：
Hu, Jingzhou
影响因子：
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作者：
Beniaminov AD;Puzanov GA;Krasnov GS;Kaluzhny DN;Kazubskaya TP;Braga EA;Kudryavtseva AV;Melnikova NV;Dmitriev AA
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Dmitriev AA
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20.8
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通讯作者：
Rabinowitz, Joshua D.
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2.9
作者：
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