MOLECULAR & CELLULAR PROTEOMICS
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2018
·Vol. 17
·期 10
·2005-2017
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IF 5.236
Weixuan Wang,Yadong Hu,Changmei Yang,Songbiao Zhu,Xiaofei Wang,Zhenyu Zhang,Haiteng Deng
Nicotinamide adenine dinucleotide (NAD) plays an essential role in all aspects of human life. NAD levels decrease as humans age, and supplementation with NAD precursors plays a protective role against aging and associated disease. Less is known about the effects of decreased NAD on cellular processes, which is the basis for understanding the relationship between cellular NAD levels and aging-associated disease. In the present study, cellular NAD levels were decreased by overexpression of CD38, a NAD hydrolase, or by treating cells with FK866, an inhibitor of nicotinamide phosphoribosyltransferase (NAMPT). Quantitative proteomics revealed that declining NAD levels downregulated proteins associated with primary metabolism and suppressed cell growth in culture and nude mice. Decreased glutathione synthesis caused a 4-fold increase in cellular reactive oxygen species levels, and more importantly upregulated proteins related to movement and adhesion. In turn, this significantly changed cell morphology and caused cells to undergo epithelial to mesenchymal transition (EMT). Secretomic analysis also showed that decreased NAD triggered interleukin-6 and transforming growth factor beta (TGFβ) secretion, which activated integrin-β-catenin, TGFβ-MAPK, and inflammation signaling pathways to sustain the signaling required for EMT. We further revealed that decreased NAD inactivated sirtuin 1, resulting in increased signal transducer and activator of transcription 3 (STAT3) acetylation and phosphorylation, and STAT3 activation. Repletion of nicotinamide or nicotinic acid inactivated STAT3 and reversed EMT, as did STAT3 inhibition. Taken together, these results indicate that decreased NAD activates multiple signaling pathways to promote EMT and suggests that age-dependent decreases in NAD may contribute to tumor progression. Consequently, repletion of NAD precursors has potential benefits for inhibiting cancer progression.
研究领域:
细胞生物学
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分子生物学
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癌症研究
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衰老研究
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产品关联
DOI
PubMed
高影响因子
PHYTOMEDICINE
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2024
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IF 6.7
Wenjuan Liu,Junjie Li,Sihan Xu,Yixu Wang,Jiawei Li,Shou Wang,Li Fu,Min Jiang,Gang Bai
ABSTRACT Background Fever is one of the main pathophysiological reactions that occurs during the acute phase of various diseases. Excessive body temperature can lead to various adverse consequences such as brain tissue damage and abnormal immune responses. Phillyrin (Phr) is the main active ingredient in Forsythia suspensa (Thunb.) Vahl (Lian Qiao) and has antipyretic effects; however, its antipyretic mechanism of action remains unclear. Purpose This study aimed to explore the antipyretic mechanisms of Phr and provide a new treatment plan for fever. Methods The antipyretic effects of Phr were evaluated using a mouse model of pneumonia fever. The main metabolites of Phr involved in its antipyretic function were identified using a mitochondrial temperature-sensitive probe. Further synthesis of the main metabolite, phillygenin (Phg), an alkynylated probe, was performed, and chemical proteomics was used to capture and analyze its direct target for antipyretic effects. The mechanism of action of Phg and its antipyretic targets was explored using metabolomics and various molecular biology methods. Results Phr showed significant antipyretic and anti-inflammatory effects in a mouse model of lipopolysaccharide-induced fever. Phg reversibly targeted the nicotinamide adenine dinucleotide (NAD + ) binding domain of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), malate dehydrogenase 2 (MDH2), and isocitrate dehydrogenase 2 (IDH2) to inhibit their enzymatic activity. In-depth analysis of cellular metabolomics and mitochondrial stress testing indicated that inhibition of GAPDH, MDH2, and IDH2 enzyme activity by Phg led to a decrease in cellular energy supply and heat production regulated by glycolysis, tricarboxylic acid cycle, and oxidative phosphorylation signaling pathways. Phg specifically targeted macrophages and inhibited LPS-induced macrophage activation by downregulating GAPDH enzyme activity, thereby exerting anti-inflammatory effects. In vivo experiments also confirmed that the antipyretic effect of Phr in LPS-induced fever model mice was related to its main metabolites, Phg and Phg-sulfonate (Phg-S), which directly targeted the NAD + binding domain of GAPDH, IDH2, and MDH2, inhibiting the activity of these enzymes, thereby reducing energy supply and regulating febrile-related inflammatory factors. Conclusion This study reported for the first time that the antipyretic effect of Phr is produced by targeting GAPDH, IDH2, and MDH2 to regulate energy supply and febrile-related inflammatory factors through its main metabolites Phg and Phg-S. This study not only provides potential drugs for fever treatment but also provides new ideas for improving clinical fever treatment plans.
研究领域:
药理学
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生物化学
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分子生物学
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免疫学
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DOI
PubMed
高影响因子
CANCER LETTERS
·
2024
·
IF 9.1
Huimin Huang,Shitong Wang,Hongping Xia,Xingling Zhao,Kaiyuan Chen,Guihua Jin,Shipeng Zhou,Zhaoliang Lu,Tongke Chen,Huajun Yu,Xiaoqun Zheng,Haishan Huang,Linhua Lan
The aim of this study was to investigate the underlying molecular mechanism behind the promotion of cell survival under conditions of glucose deprivation by l -lactate. To accomplish this, we performed tissue microarray and immunohistochemistry staining to analyze the correlation between the abundance of pan-Lysine lactylation and prognosis. In vivo evaluations of tumor growth were conducted using the KPC and nude mice xenograft tumor model. For mechanistic studies, multi-omics analysis, RNA interference, and site-directed mutagenesis techniques were utilized. Our findings robustly confirmed that l -lactate promotes cell survival under glucose deprivation conditions, primarily by relying on GLS1-mediated glutaminolysis to support mitochondrial respiration. Mechanistically, we discovered that l -lactate enhances the NMNAT1-mediated NAD + salvage pathway while concurrently inactivating p-38 MAPK signaling and suppressing DDIT3 transcription. Notably, Pan-Kla abundance was significantly upregulated in patients with Pancreatic adenocarcinoma (PAAD) and associated with poor prognosis. We identified the 128th Lysine residue of NMNAT1 as a critical site for lactylation and revealed EP300 as a key lactyltransferase responsible for catalyzing lactylation. Importantly, we elucidated that lactylation of NMNAT1 enhances its nuclear localization and maintains enzymatic activity, thereby supporting the nuclear NAD + salvage pathway and facilitating cancer growth. Finally, we demonstrated that the NMNAT1-dependent NAD + salvage pathway promotes cell survival under glucose deprivation conditions and is reliant on the activity of Sirt1. Collectively, our study has unraveled a novel molecular mechanism by which l -lactate promotes cell survival under glucose deprivation conditions, presenting a promising strategy for targeting lactate and NAD + metabolism in the treatment of PAAD.
研究领域:
癌症生物学
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代谢
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分子生物学
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细胞生物学
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DOI
PubMed
高影响因子
MOLECULAR MEDICINE
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2024
·Vol. 30
·期 1
·1-14
·
IF 6
Yu Hao,Gan Daojing,Luo Zhen,Yang Qilin,An Dongqi,Zhang Hao,Hu Yingchun,Ma Zhuang,Zeng Qingchun,Xu Dingli,Ren Hao
Background In heart failure (HF), mitochondrial dysfunction and metabolic remodeling lead to a reduction in energy productivity and aggravate cardiomyocyte injury. Supplementation with α-ketoglutarate (AKG) alleviated myocardial hypertrophy and fibrosis in mice with HF and improved cardiac insufficiency. However, the myocardial protective mechanism of AKG remains unclear. We verified the hypothesis that AKG improves mitochondrial function by upregulating NAD + levels and activating silent information regulator 2 homolog 1 (SIRT1) in cardiomyocytes. Methods In vivo, 2% AKG was added to the drinking water of mice undergoing transverse aortic constriction (TAC) surgery. Echocardiography and biopsy were performed to evaluate cardiac function and pathological changes. Myocardial metabolomics was analyzed by liquid chromatography‒mass spectrometry (LC‒MS/MS) at 8 weeks after surgery. In vitro, the expression of SIRT1 or PINK1 proteins was inhibited by selective inhibitors and siRNA in cardiomyocytes stimulated with angiotensin II (AngII) and AKG. NAD + levels were detected using an NAD test kit. Mitophagy and ferroptosis levels were evaluated by Western blotting, qPCR, JC-1 staining and lipid peroxidation analysis. Results AKG supplementation after TAC surgery could alleviate myocardial hypertrophy and fibrosis and improve cardiac function in mice. Metabolites of the malate-aspartate shuttle (MAS) were increased, but the TCA cycle and fatty acid metabolism pathway could be inhibited in the myocardium of TAC mice after AKG supplementation. Decreased NAD + levels and SIRT1 protein expression were observed in heart of mice and AngII-treated cardiomyocytes. After AKG treatment, these changes were reversed, and increased mitophagy, inhibited ferroptosis, and alleviated damage in cardiomyocytes were observed. When the expression of SIRT1 was inhibited by a selective inhibitor and siRNA, the protective effect of AKG was suppressed. Conclusion Supplementation with AKG can improve myocardial hypertrophy, fibrosis and chronic cardiac insufficiency caused by pressure overload. By increasing the level of NAD + , the SIRT-PINK1 and SIRT1-GPX4 signaling pathways are activated to promote mitophagy and inhibit ferroptosis in cardiomyocytes, which ultimately alleviates cardiomyocyte damage.
研究领域:
心脏病学
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分子生物学
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代谢学
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药理学
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DOI
PubMed
高影响因子
ADVANCED SYNTHESIS & CATALYSIS
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2022
·Vol. 364
·期 8
·1450-1459
·
IF 5.981
Feng Cheng,Jia-Min Zhang,Zhen-Tao Jiang,Xiao-Hu Wu,Ya-Ping Xue,Yu-Guo Zheng
Chiral amino acids are extensively applied in the pharmaceutical, food, cosmetic, and agricultural industries. As a representative example, l -phosphinothricin ( l -PPT, a chiral non-natural amino acid) is a broad-spectrum herbicide. An NAD(H)-driven biocatalytic system for the asymmetric synthesis of chiral amino acids (focused on l -PPT) with high efficiency and low cost is highly desired. The key for the development of such biocatalytic system is to obtain an NADH-dependent biocatalyst with high catalytic performance toward l -PPT pro-ketone PPO. Herein, an engineered glutamate dehydrogenase from Lysinibacillus composti ( Lc GluDH) with desired activity was obtained by gene mining and protein engineering. In silico analyses suggested that the volume of substrate-binding pocket was substantially enlarged from 330.5 Å 3 to 409.6 Å 3 . The stability of Lc GluDH was increased ( T m value increased from 47.3 °C to 65.3 °C) by introducing positively charged amino acids or aromatic amino acids at position 375. The desired biocatalytic system was constructed by coupling the engineered Lc GluDH and an NAD + -dependent FDH. Through this biocatalytic system, the batch production of l -PPT exhibited high space-time yield (207.3 g ⋅ L −1 ⋅ day −1 ) with strict stereoselectivity ( ee of l -PPT>99%). Furthermore, eight other chiral amino acids were synthesised by the developed NAD(H)-driven biocatalytic system with high ee values.
研究领域:
生物化学
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生物技术
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药物化学
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酶学
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产品关联
DOI
高影响因子
ADVANCED MATERIALS
·
2024
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IF 27.4
Ying Kong,Xu Chen,Feng Liu,Jiageng Tang,Yijing Zhang,Xiangxiang Zhang,Luyao Zhang,Tong Zhang,Yaqi Wang,Mengxiao Su,Qixin Zhang,Hanxiang Chen,Di Zhou,Fan Yi,Hong Liu,Yi Fu
As a central metabolic molecule, nicotinamide adenine dinucleotide (NAD + ) can potentially treat acute kidney injury (AKI) and chronic kidney disease (CKD); however, its bioavailability is poor due to short half-life, instability, the deficiency of targeting, and difficulties in transmembrane transport. Here we designed a physiologically adaptive gallic acid-NAD + nanoparticle, that had ultrasmall size and pH-responsiveness, passed through the glomerular filtration membrane to reach injured renal tubules, and efficiently delivered NAD + into the kidneys. With an effective accumulation in the kidneys, it restored renal function, immune microenvironment homeostasis, and mitochondrial homeostasis of AKI mice via the NAD + -Sirt1 axis, and exerted strong antifibrotic effects on the AKI-to-CKD transition by inhibiting TGF-β signaling. It also exhibited excellent stability, biodegradable, and biocompatible properties, ensuring its long-term safety, practicality, and clinical translational feasibility. The present study shows a potential modality of mitochondrial repair and immunomodulation through nanoagents for the efficient and safe treatment of AKI and CKD. This article is protected by copyright. All rights reserved
研究领域:
肾病学
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药学
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生物医学工程
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分子生物学
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DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
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2024
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IF 3.7
Shu-Ping Zou,Wei Ding,Yu-Yue Han,Kun Niu,Ya-Ping Xue,Yu-Guo Zheng
L-Phosphinothricin (L-PPT) is the active compound of a broad-spectrum herbicide. In this study, we biosynthesized L-PPT via the asymmetric reduction of 2-carboxy-4-(hydroxymethylphosphoryl) butyric acid (PPO) using a whole-cell biocatalyst. Glutamate dehydrogenase from Pseudomonas moorei ( Pm GluDH), Glucose dehydrogenase from Exiguobacterium sibiricum ( Es GDH) and NAD kinase from Corynebacterium glutamicum ( Cg NADK) were coexpressed in E. coli , significantly increasing the catalytic efficiency by regenerating more NADPH to promote PPO reduction. Following the optimization of balancing the co-expression of the three enzymes in pETDuet-1 and pCDFDuet-1, the coexpressing strain increases in the intracellular NADP(H) concentration to 54.9 μmol/g DCW, which was 3.8-fold higher than the control. The co-expression system exhibited optimal activity at a temperature of 35°C, pH 7.5, and the glucose to PPO molar ratio of 1.4:1. In a 30 L bioconversion system the final concentration and productivity of L-PPT reached 79.8 g/L (>99% ee ) and 1440 g/(L·d), respectively. These results will contribute to the industrial biological production of L-PPT.
研究领域:
生物技术
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生化工程
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代谢工程
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产品关联
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
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2021
·
IF 10.618
Haiyan Song,Xigui Zhou,Zhiguang Zhu
NAD + -dependent dehydrogenase-based biosensors usually suffer from the low accuracy due to the interference of cofactors in the complex environment, such as fermentation samples. Herein, we demonstrate the example of an integrated biosensor device that can be applied for analyzing xylose fermentation samples. The device is composed of one chamber for the elimination of NAD + and NADH in the fermentation broth and another chamber for the sample analysis. In the first chamber, a cyclic voltammetry method coupled with Ni foam as a working electrode was proven to be effective in removing NAD + and NADH in the fermentation broth. In the other chamber, xylose dehydrogenase , as the recognition element, and diaphorase, used for the regeneration of bioactive NAD + mediated by vitamin K 3 , were co-immobilized on the surface of the magnetic nanoparticles , which was further coated onto a magnetic glassy carbon electrode. The detection range of the constructed biosensor was from 0.5 to 10 g L −1 with a detection limit of 0.01 g L −1 at a signal-to-noise ratio of 3. Moreover, the biosensor achieved high selectivity, recovery, reproducibility, and good long-time stability when analyzing real xylose fermentation samples, suggesting its promising application potential.
研究领域:
生物技术
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分析化学
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电化学
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发酵技术
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DOI
高影响因子
FITOTERAPIA
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2024
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IF 2.5
Liwenyu Chen,Yi Yu,Min Lin,Ming Liang,Chen Yang,Xutian Kan,Xiaoliang Lin,Jin Qi
Schisandra chinensis (Turcz.) Baill, a traditional Chinese medicine with significant nourishing functions, has a regulatory effect on the cardiovascular system, digestive system, central nervous system, and endocrine and immune systems. It can protect the cardiovascular system, improve immunity, and has anti-oxidant and anti-aging properties. This study aimed to identify the chisandra chinensis components that increased NAD+ levels, by using spectrum-effect analysis and experimental validations. First, the quality of the S. chinensis extract was analyzed by HPLC-MS. The extract of S. chinensis increased the NAD+ levels of HUVECs during oxygen and glucose deprivation injury, and protected HUVEC s from injury from superoxide dismutase (SOD) and aging. The relationships of spectral effects were studied by partial least square regression. Coupled with target cell extraction, the material basis for increasing NAD+ levels in S. chinensis was obtained. The pharmacological activity of S. chinensis was verified at the cellular level. The related enzymes of the NAD+ synthesis and decomposition pathways were identified, showing that S. chinensis increased the level of NAD+ by increasing the activity of related enzymes in the synthesis pathways of NMNAT1, NMNAT2, NMNAT3, and NAMPT, but had no effect on the decomposition pathway. Finally, four constituents were confirmed, in vitro, to be the basis of S. chinensis -induced increases in the levels of NAD+. The EC 50 values of the four constituents were also determined.
研究领域:
药理学
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心血管生物学
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植物化学
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产品关联
DOI
PubMed
高影响因子
Mucosal Immunology
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2024
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IF 7.9
Jinzhe Tong,Simiao Chen,Xinyue Gu,Xuanqi Zhang,Fang Wei,Yingying Xing
Tissue-resident memory T cells (T RM ) can be induced by infection and vaccination, and play a key role in maintaining long-term protective immunity against mucosal pathogens. Our studies explored the key factors and mechanisms affecting the differentiation, maturation, and stable residence of gastric epithelial CD4 + T RM induced by Helicobacter pylori (Hp) vaccine and optimized Hp vaccination to promote the generation and residence of T RM .CD38 regulated mitochondrial activity and enhanced TGF-β signal transduction to promote the differentiation and residence of gastric epithelial CD4 + T RM by mediating the expression of CD105. Extracellular nucleotides influenced the long-term maintenance of T RM in gastric epithelium by P2RX7. Vitamin D3 and Gram-positive enhancer matrix particles (GEMs)as immune adjuvants combined with Hp vaccination promoted the production of CD69 + CD103 + CD4 + T RM .
研究领域:
免疫学
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微生物学
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分子生物学
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疫苗学
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DOI
PubMed
高影响因子
Advanced Science
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2025
·
IF 14.1
Shuoting Zhou,Xi Xiong,Jialong Hou,Qi Duan,Yi Zheng,Tao Jiang,Jiani Huang,Haijun He,Jiaxue Xu,Keke Chen,Wenwen Wang,Jinlai Cai,Jingjing Qian,Huijun Chen,Weihong Song,XinShi Wang,Chenglong Xie
Serving as a pivotal hub for cellular metabolism and intracellular signaling, the mitochondrion has emerged as a crucial organelle whose dysfunction is linked to many human diseases, including neurodegenerative disorders, particularly Parkinson's disease (PD). However, whether mitochondrial quality control (MQC) can be targeted for therapeutic interventions remains uncertain. This study uses clinical samples, molecular biology techniques, pharmacological interventions, and genetic approaches to investigate the significance of NAD+ levels in cross-species models of PD. These results reveal that treatment of rotenone-incubated cells with NAD+ boosters (such as NMN, siCD38, and NAT) increases UPR mt /mitophagy-related MQC, reduces pro-inflammatory cytokine expression, inhibits apoptosis, and strengthen redox reactions. In vivo, NMN supplementation inhibits motor deficit and forestalls the neuropathological phenotypes of MPTP-induced PD mice, which are required for the atf4-related mitochondrial UPR pathway. Notably, bulk omics signatures and metabolomic profiling analyses of the striatum reveal NMN-induced transcriptional changes in genes and proteins involved in mitochondrial homeostasis. Thus, these findings demonstrate that the accelerated pathology in PD models is probably mediated by impaired MQC and that bolstering cellular NAD+ levels alleviates mitochondrial proteotoxic stress and mitigate PD phenotypes.
研究领域:
神经科学
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细胞生物学
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药理学
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分子生物学
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生物化学
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基因组学
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代谢组学
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产品关联
DOI
PubMed
高影响因子
Nature Communications
·
2023
·Vol. 14
·期 1
·1-12
·
IF 16.6
Liu Jianming,Zhang Han,Xu Yingying,Meng Hao,Zeng An-Ping
The use of gaseous and air-captured CO 2 for technical biosynthesis is highly desired, but elusive so far due to several obstacles including high energy (ATP, NADPH) demand, low thermodynamic driving force and limited biosynthesis rate. Here, we present an ATP and NAD(P)H-free chemoenzymatic system for amino acid and pyruvate biosynthesis by coupling methanol with CO 2 . It relies on a re-engineered glycine cleavage system with the NAD(P)H-dependent L protein replaced by biocompatible chemical reduction of protein H with dithiothreitol. The latter provides a higher thermodynamic driving force, determines the reaction direction, and avoids protein polymerization of the rate-limiting enzyme carboxylase. Engineering of H protein to effectively release the lipoamide arm from a protected state further enhanced the system performance, achieving the synthesis of glycine, serine and pyruvate at g/L level from methanol and air-captured CO 2 . This work opens up the door for biosynthesis of amino acids and derived products from air.
研究领域:
生物技术
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化学工程
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生物化学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）