Quantitative Proteomic Approach Reveals Altered Metabolic Pathways in Response to the Inhibition of Lysine Deacetylases in A549 Cells under Normoxia and Hypoxia

Growing evidence is showing that acetylation plays an essential role in cancer, but studies on the impact of KDAC inhibition (KDACi) on the metabolic profile are still in their infancy. Here, we analyzed, by using an iTRAQ-based quantitative proteomics approach, the changes in the proteome of KRAS-mutated non-small cell lung cancer (NSCLC) A549 cells in response to trichostatin-A (TSA) and nicotinamide (NAM) under normoxia and hypoxia. Part of this response was further validated by molecular and biochemical analyses and correlated with the proliferation rates, apoptotic cell death, and activation of ROS scavenging mechanisms in opposition to the ROS production. Despite the differences among the KDAC inhibitors, up-regulation of glycolysis, TCA cycle, oxidative phosphorylation and fatty acid synthesis emerged as a common metabolic response underlying KDACi. We also observed that some of the KDACi effects at metabolic levels are enhanced under hypoxia. Furthermore, we used a drug repositioning machine learning approach to list candidate metabolic therapeutic agents for KRAS mutated NSCLC. Together, these results allow us to better understand the metabolic regulations underlying KDACi in NSCLC, taking into account the microenvironment of tumors related to hypoxia, and bring new insights for the future rational design of new therapies.     

Xylose-lysine model systems: The effect of pH on the volatile reaction products

Aqueous molal solutions of xylose and lysine (initial pH 4–9) were refluxed either with control of the pH at 5–0 or without pH control (final pH 2–6). Analysis by gas chromatography (GC) and GC-mass spectrometry resulted in the identification of 58 and 28 compounds, respectively, from the two systems. Furans were the main reaction products in both systems and 2-furfural alone comprised 522 and 999 g kg^?1 of the volatiles, respectively, from the systems with final pH values of 5–0 and 2–6. Maintaining the pH at 5–0 resulted in a higher yield and greater numbers of nitrogen-containing compounds, and monocyclic pyrroles, pyridines and 2,3-dihydro-l H-pyrrolizines were identified only in that system. Aliphatic compounds, alicyclic compounds, benzene derivatives. l-(2–furfuryl)pyrroles and pyrazines were also identified. This investigation is the first report of the formation of 2.3-dihydro-l H-pyrrolizines in a model system containing lysine as the amino compound; a possible mechanism is proposed.     

Evidence for a Negative Correlation between Human Reactive Enamine-Imine Intermediate Deaminase A (RIDA) Activity and Cell Proliferation Rate: Role of Lysine Succinylation of RIDA

Reactive intermediate deaminase (Rid) proteins are enzymes conserved in all domains of life. UK114, a mammalian member of RidA subfamily, has been firstly identified as a component of liver perchloric acid-soluble proteins (L-PSP). Although still poorly defined, several functions have been attributed to the mammalian protein UK114/RIDA, including the reactive intermediate deamination activity. The expression of UK114/RIDA has been observed in some tumors, arousing interest in this protein as an evaluable tumor marker. However, other studies reported a negative correlation between UK114/RIDA expression, tumor differentiation degree and cell proliferation. This work addressed the question of UK114/RIDA expression in human non-tumor HEK293 cell lines and in some human tumor cell lines. Here we reported that human RIDA (hRIDA) was expressed in all the analyzed cell line and subjected to lysine (K-)succinylation. In HEK293, hRIDA K-succinylation was negatively correlated to the cell proliferation rate and was under the control of SIRT5. Moreover, K-succinylation clearly altered hRIDA quantification by immunoblotting, explaining, at least in part, some discrepancies about RIDA expression reported in previous studies. We found that hRIDA was able to deaminate reactive enamine-imine intermediates and that K-succinylation drastically reduced deaminase activity. As predicted by in silico analysis, the observed reduction of deaminase activity has been related to the drastic alterations of hRIDA structure inferred by K-succinylation. The role of hRIDA and the importance of its K-succinylation in cell metabolism, especially in cancer biology, have been discussed.     

The Mystery of Extramitochondrial Proteins Lysine Succinylation

Lysine succinylation is a post-translational modification which alters protein function in both physiological and pathological processes. Mindful that it requires succinyl-CoA, a metabolite formed within the mitochondrial matrix that cannot permeate the inner mitochondrial membrane, the question arises as to how there can be succinylation of proteins outside mitochondria. The present mini-review examines pathways participating in peroxisomal fatty acid oxidation that lead to succinyl-CoA production, potentially supporting succinylation of extramitochondrial proteins. Furthermore, the influence of the mitochondrial status on cytosolic NAD⁺ availability affecting the activity of cytosolic SIRT5 iso1 and iso4—in turn regulating cytosolic protein lysine succinylations—is presented. Finally, the discovery that glia in the adult human brain lack subunits of both alpha-ketoglutarate dehydrogenase complex and succinate-CoA ligase—thus being unable to produce succinyl-CoA in the matrix—and yet exhibit robust pancellular lysine succinylation, is highlighted.     

全球氨基酸市场供需与预测

概述了近年来饲料添加剂用氨基酸类的现状和今后发展。着重介绍了氨基酸中主要产品蛋氨酸、赖氨酸和苏氨酸在美国、欧盟和日本的生产和市场发展动向。由于这类食品添加剂需求强劲，目前，该市场正处在回升之中。     

L-赖氨酸铜的制备及工艺优化

为解决氨基酸金属螯合物采用NaOH调解体系pH值时其反应溶液碱性不易控制的问题,以L-赖氨酸和氯化铜为原料,选用NH3.H2O作为体系pH调节剂,合成了L-赖氨酸铜。单因素平行试验优选法得到L-赖氨酸铜的合成工艺条件为：氯化铜与赖氨酸盐酸盐的摩尔比为3∶5,pH≈8,反应温度为45℃,反应时间为60 min。反应全程在液相中进行,条件温和,避免了因碱性过大而出现沉淀;产品收率为71%;合成的L-赖氨酸铜为蓝色粉末,易溶于水,不溶于乙醇、乙醚等有机溶剂,熔点为227.3～227.5℃。     

赖氨酸生理功能的研究进展

赖氨酸具有促进机体生长发育、参与能量代谢、促进矿物质吸收和骨骼生长、增强免疫功能、治疗单纯疱疹病毒感染、减轻焦虑等功能,综述与赖氨酸生理功能有关的研究进展表明：赖氨酸不仅可作为营养强化剂促进身体健康,而且可作为某些药物的替代品用于疾病的预防和治疗,以减少药物的副作用。     

赖氨酸生产与应用发展概况

目前 ,我国的赖氨酸生产能力超过 10万t/a,实际生产量只有 3万t/a。主要原因是我国的赖氨酸生产不稳定 ,产品成本高 ,品种单一 ,使得进口产品不断冲击国内市场 ,以致于国内现有的生产装置不能满负荷运转。近几年 ,国内赖氨酸新建和扩建装置不断增多 ,但进口产品量也不断增加 ,这一点非常值得注意。