CURRENT ISSUES IN MOLECULAR BIOLOGY
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2026
·Vol. 48
·期 5
·472
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IF 4.1
Lina Wang,Lian Wu,Kexin Zhang,Rui Ma,Shurong Chen,Tong Ji,Min Zhou,Jiayi Xie,Lingli Zheng,Qingshan Bill Fu
Brucellosis, acting as a typical chronic zoonotic disease, is caused by the invasion ofBrucellainto the human body. Outer membrane protein 25 (Omp25), specifically localized on theBrucellamembrane, is the main virulence factor ofBrucellaand participates in multiple links of the damage process. Omp25c, a porin protein ofBrucella, is a paralog of Omp25 with high sequence identity. NADH dehydrogenase [ubiquinone] complex I assembly factor 2 (Ndufaf2) has a key function in cell energy metabolism, particularly in the formation and activity of the mitochondrial respiratory chain. Loss of Ndufaf2 results in oxidative stress and mitochondrial DNA (mtDNA) deletion. However, the functional relationship between Omp25c and Ndufaf2, the underlying mechanism of the proteins, remains unclear. In this work, we purified the Omp25c and Ndufaf2proteins. Our data revealed that Omp25c directly interacts with Ndufaf2, as determined using Biacore analysis. In addition, assays revealed that Ompa2c reshapes the host cell’s redox environment by decreasing the oxidized nicotinamide adenine dinucleotide/reduced nicotinamide adenine dinucleotide (NAD+/NADH) ratioand adenosine triphosphate (ATP) production, whereas Ndufaf2 exerts an opposing regulatory effect; Co-expression results further revealed an antagonistic relationship between the two during metabolic processes. These findings provide a new perspective for elucidating the mechanisms of mitochondrial functional regulation inBrucella–host interactions and lay the theoretical and experimental foundation for drug development targeting metabolic interventions to eliminate intracellular pathogens.
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BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
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2024
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IF 2.5
Xuanming Chang,Xiaoqian Chen,Xiuze Zhang,Nian Chen,Weitao Tang,Zhuo Zhang,Sulin Zheng,Jiong Huang,Yihan Ji,Yuzheng Zhao,Yi Yang,Xie Li
Lactate plays a crucial role in energy metabolism and greatly impacts protein activities, exerting diverse physiological and pathological effects. Therefore, convenient lactate assays for tracking spatiotemporal dynamics in living cells are desirable. In this paper, we engineered and optimized a red fluorescent protein sensor for l -lactate named FiLa-Red. This indicator exhibited a maximal fluorescence change of 730 % and an apparent dissociation constant ( K d ) of approximately 460 μM. By utilizing FiLa-Red and other sensors, we monitored energy metabolism in a multiplex manner by simultaneously tracking lactate and NAD + /NADH abundance in the cytoplasm, nucleus, and mitochondria. The FiLa-Red sensor is expected to be a useful tool for performing metabolic analysis in vitro , in living cells and in vivo .
研究领域:
细胞生物学
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分子生物学
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生物技术
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代谢
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CARBON
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2025
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IF 11.6
Junzhu Yang,Junchen Liu,Yi Zhang,Wei Pei,Qi Xia,Sanyang Han,Yuan Lu
Efficient regeneration of the coenzymes NADH and NADPH is crucial for sustaining biocatalytic processes, particularly in driving the biocatalysis reaction. In this study, the photocatalytic properties of alkali-doped graphitic carbon nitrates were explored to achieve efficient NAD(P)H regeneration. g-C 3 N 4 doped with potassium (KOH), sodium (NaOH), and calcium (Ca(OH) 2 ) was synthesized and characterized, showing significant enhancements in light absorption, charge separation, and photogenerated electron transport. Among the synthesized photocatalysts, KOH-doped g-C 3 N 4 (KCN) exhibited the best photocatalytic performance, regenerating 95 % NADH and 92 % NADPH within 30 min. This result compares favorably with those reported in previous studies. Based on this superior performance, K-0.2 was further applied to the in vitro CO 2 reduction to formate as a proof of concept. To clarify the mechanistic insights of the improved photocatalytic performance of KCN, the chemical model of g-C 3 N 4 and KCN was established, and DFT calculations were performed to reveal the electronic structure modifications responsible for the enhanced photocatalytic activity. Furthermore, to expand the light utilization capability of the system, upconversion nanoparticles (UCNPs) were incorporated into the system to utilize near-infrared light (980 nm), which successfully achieved NADH regeneration under near-infrared excitation and extended the light absorption range of the photocatalytic system. Overall, this study demonstrated that alkali-doped g-C 3 N 4 offered a highly efficient platform for NAD(P)H regeneration, and the integration of UCNPs further enriched the coenzyme regeneration scenario, suggesting a promising strategy for advancing sustainable coenzyme regeneration technologies.
研究领域:
生物催化
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光化学
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分子生物学
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可持续能源
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酶工程
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2019
·
IF 4.784
Meng-Qiu Xu,Fei-Long Li,Wen-Qian Yu,Rui-Fang Li,Ye-Wang Zhang
Cofactor regeneration is an important method to avoid the consumption of large quantities of oxidized cofactor NAD + in enzyme-catalyzed reactions. Herein, glycerol dehydrogenase (GDH) and NADH oxidase preparations by aggregating enzymes with ammonium sulphate followed by cross-linking formed aggregates for effective regeneration of NAD + . After optimization, the activity of combi-CLEAs and separate CLEAs mixtures were 950 and 580 U/g, respectively. And the catalytic stability of combi-CLEAs against pH and temperature was superior to the free enzyme mixture. After ten cycles of reuse, the catalytic efficiency could still retain 63.3% of its initial activity, indicating that the constructed combi-CLEAs system had excellent reusability. Also, the conversion of glycerol to 1,3-dihydroxyacetone (DHA) was improved by the constructed NAD + regeneration system, resulting in 4.6%, which was 2.5 times of the free enzyme system. Thus, wide applications of this co-immobilization method in the production of various chiral chemicals could be expected in the industry for its high efficiency at a low cost.
研究领域:
生物化学
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酶工程
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NUCLEIC ACIDS RESEARCH
·
2026
·Vol. 54
·期 5
·
IF 15
Liu Wuzhen,Hu Kefan,Nie Shiqi,Feng Zhang,Li Qiongfang,Zhang Hailei,Liang Shumin,Peng Jialin,Zhang Shoudong,Cai Zongwei,Xia Yiji
Recently, nicotinamide adenine dinucleotide (NAD+) and other nucleotide analogs have been identified as non-canonical RNA caps in both prokaryotes and eukaryotes. In Escherichia coli, NAD capping has been shown to be influenced by environmental conditions in a gene-specific manner, yet its regulatory mechanisms remain poorly understood. We previously reported that most transcripts produced by sibD are NAD-capped during the stationary phase. In this study, we found that the 35-bp minimal promoter of sibD is sufficient for its NAD capping. When this minimal promoter was applied to express genes not typically producing NAD-RNAs, their transcripts could also be NAD-capped. These findings strongly support that NAD capping in E. coli occurs during transcription initiation mediated by the promoter and RNA polymerase. Additionally, the bacterial alarmone ppGpp and the small protein DksA, both transcription initiation regulators, synergistically enhance transcription of both NAD-capped and uncapped RNAs from sibD and its homologous genes, sibC and sibE, in vitro. In contrast, the ppGpp⁰ mutant, deficient in ppGpp synthesis, showed a significant reduction in NAD-RNA production from these genes. Our findings elucidate the cis-elements and trans-acting factors mediating NAD capping during transcription initiation, offering novel insight into this regulatory process.
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ADVANCED MATERIALS
·
2026
·
IF 29.1
Jingtao Liu,Xun Guo,Nan Wang,Xingyu Zhang,Xiaoting Wang,Guangru Li,Xumin Hou,Luodan Yu,Yu Chen,Ruogu Li
Myocardial ischemia/reperfusion (I/R) injury is a globally recognized pathological condition that results in life-threatening organ dysfunction. Supplementation with nicotinamide adenine dinucleotide (NAD + ) can improve myocardial energy metabolism, alleviate inflammation, and mitigate I/R injury. However, its clinical applicability is constrained by the poor efficacy and delivery limitations of current NAD + supplementation strategies. Here, we design a dual single-atom nanozyme with nicotinamide adenine dinucleotide (NADH) oxidase-mimicking activity for in situ NAD + biotransformation and I/R injury treatment. Upon M2 macrophage membrane coating, the nanozyme selectively targets ischemic myocardium, facilitating in situ NAD + biotransformation by eliminating excess NADH, while concurrently scavenging reactive oxygen species, restoring mitochondrial bioenergetics, attenuating inflammation, and inhibiting apoptosis. Mechanistic investigations demonstrate that the nanozyme activates the NAD + -SIRT1 axis, enhancing ATP biosynthesis and oxidative phosphorylation, while suppressing TNF signaling and NF-κB pathways. Collectively, the engineered NOX-like nanozyme offers a novel therapeutic strategy for myocardial I/R injury by modulating both mitochondrial bioenergetics and oxidative stress defense pathways through restoring NAD + /NADH homeostasis.
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PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR
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2025
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IF 2.5
Yikai Chen,Wen Ye,Wenxin Wang,Nan Miao,Yongqiang Sha,Tao Sun
Major depressive disorder (MDD) is characterized by anhedonia and cognitive deficits. In this study, we investigated the therapeutic effects of nicotinic acid (NA), a nicotinamide adenine dinucleotide + (NAD + ) precursor, in a mouse chronic restraint stress (CRS) model. Behavioral assessments revealed that NA ameliorated depressive-like behaviors, evidenced by restored sucrose preference and reduced immobility in forced swim and tail suspension tests, without affecting motor coordination in the rotarod test. Hippocampal transcriptomic analysis indicated that NA reversed stress-induced gene dysregulation, activated neuroprotective Wnt/β-catenin, cGMP–PKG, and cAMP pathways, and rescued lipid and porphyrin metabolism. In conclusion, NA likely exerts antidepressant effects by remodeling hippocampal gene networks to enhance synaptic plasticity and restore metabolic homeostasis. These findings highlight NA as a promising metabolic antidepressant and support the development of NAD + precursor–based potential therapies for mood disorders.
研究领域:
神经生物学
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精神病学
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分子药理学
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代谢组学
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转化医学
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表观遗传学
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神经内分泌学
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JOURNAL OF PHOTOCHEMISTRY AND PHOTOBIOLOGY A-CHEMISTRY
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2022
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IF 5.141
Shengnan Fan,Xiaoqing Li,Fanghui Ma,Minghui Yang,Juan Su,Xiang Chen
Lactate dehydrogenase (LDH) plays an important role in the process of glycolysis as well as gluconeogenesis, and is related to many diseases, so it is of great significance to accurately detect LDH activity in serum. The fluorescence of sulfur quantum dots (SQDs) synthesized by the microwave method can be specifically enhanced by nicotinamide adenine dinucleotide (NAD + ). Based on this characteristic, we reported a sensitive fluorescence assay for LDH detection. In the presence of reductive coenzyme I (NADH), LDH catalyzes the reduction of pyruvate to lactic acid, and transforms NADH to NAD + , which specifically enhances the fluorescence intensity of SQDs. Based on the fluorescence intensity increase, the assay achieved detection of LDH in the range from 500 to 40,000 U L −1 , with limit of detection of 262 U L −1 . The assay was successfully applied for the detection of LDH activity in serum, indicating potential clinical applications.
研究领域:
生物化学
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分析化学
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生物医学工程
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Nanomaterials
·
2024
·Vol. 14
·期 6
·512
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IF 4.4
Yuan Chen,Jiaxing Wang,Daidi Chen,Boxi Wang,Jinchuan Wu,Rongrong Liu,Qingxin Li
Many studies have been conducted on the microbial reduction of Pd (II) to palladium nanoparticles (Pd-NPs) due to the environmental friendliness, low cost, and the decreased toxicity of Pd (II) ions. In this study, we investigate the reduction mechanism of Pd (II) byBacillus megateriumY-4 through proteomics. The data are available via ProteomeXchange with identifier PXD049711. Our results revealed thatB. megateriumY-4 may use the endogenous electron donor (NAD(P)H) generated by nirB, tdh, and fabG and reductase to reduce Pd (II) to Pd-NPs. The expression levels of fabG, tdh, gudB, and rocG that generate NAD(P)H were further increased, and the number of reduced Pd-NPs was further increased with the exogenous electron donor sodium formate. Endogenous electron mediators such as quinones and flavins inB. megateriumY-4 can further enhance Pd (II) reduction. The findings provided invaluable information regarding the reduction mechanism of Pd (II) byB. megateriumY-4 at the proteome level.
研究领域:
微生物学
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环境科学
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NUCLEIC ACIDS RESEARCH
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2024
·
IF 16.6
Liao Fumeng,Yu Guimei,Zhang Chendi,Liu Zhikun,Li Xuzichao,He Qiuqiu,Yin Hang,Liu Xiang,Li Zhuang,Zhang Heng
Recently, a novel two-gene bacterial defense system against phages, encoding a SIR2 NADase and a HerA ATPase/helicase, has been identified. However, the molecular mechanism of the bacterial SIR2–HerA immune system remains unclear. Here, we determine the cryo-EM structures of SIR2, HerA and their complex from Paenibacillus sp. 453MF in different functional states. The SIR2 proteins oligomerize into a dodecameric ring-shaped structure consisting of two layers of interlocked hexamers, in which each subunit exhibits an auto-inhibited conformation. Distinct from the canonical AAA+ proteins, HerA hexamer alone in this antiphage system adopts a split spiral arrangement, which is stabilized by a unique C-terminal extension. SIR2 and HerA proteins assemble into a ∼1.1 MDa torch-shaped complex to fight against phage infection. Importantly, disruption of the interactions between SIR2 and HerA largely abolishes the antiphage activity. Interestingly, binding alters the oligomer state of SIR2, switching from a dodecamer to a tetradecamer state. The formation of the SIR2–HerA binary complex activates NADase and nuclease activities in SIR2 and ATPase and helicase activities in HerA. Together, our study not only provides a structural basis for the functional communications between SIR2 and HerA proteins, but also unravels a novel concerted antiviral mechanism through NAD+ degradation, ATP hydrolysis, and DNA cleavage.
研究领域:
微生物学
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结构生物学
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分子生物学
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Journal of Ovarian Research
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2026
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IF 5.3
Wang Yunduan,Li Qiao,Li Yueyue,Ma Zifeng,Xu Hongmei,Chen Kechao,Li Jian,Lan Daoliang,Fu Wei
Oxidative stress is a major driver of granulosa cell (GC) dysfunction and disrupted steroidogenesis. However, the mechanisms underlying oxidative stress–induced progesterone synthesis impairment and potential intervention strategies remain inadequately defined. In this study, RNA-seq re-analysis revealed that hydrogen peroxide (H₂O₂) markedly altered gene expression in mouse granulosa cells (mGCs), particularly affecting pathways related to progesterone biosynthesis. In vitro experiments confirmed that H₂O₂ suppresses progesterone production via downregulation of Sirtuin 1 (SIRT1). Conversely, β-nicotinamide mononucleotide (NMN) supplementation increased intracellular NAD⁺ levels, and upregulated SIRT1 and Nicotinamide phosphoribosyl transferase (NAMPT) expression. This regulation enhanced cell proliferation, stimulated the expression of progesterone biosynthetic enzymes, and promoted NAMPT transcriptional activity. Notably, the beneficial effects of NMN were diminished upon SIRT1 inhibition. Similarly, inhibition of NAMPT attenuated the positive regulatory effect of NMN, leading to a marked reduction in SIRT1 levels and a consequent suppression of progesterone secretion. Thus, SIRT1 activation induces NAMPT expression, which in turn feeds back to enhance SIRT1 activity and support progesterone biosynthesis. Together, these findings suggest that NMN alleviates H₂O₂-induced progesterone synthesis disruption in mGCs through activation of the NAD⁺/SIRT1/NAMPT axis, providing a potential therapeutic strategy for oxidative stress–related reproductive dysfunction.
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Nanomaterials
·
2021
·Vol. 11
·期 9
·2171
·
IF 5.076
Le Wang,Pengxue Sun,Yiyu Yang,Hanzhen Qiao,Hailong Tian,Dapeng Wu,Shuoye Yang,Qipeng Yuan,Jinshui Wang
The field of enzyme cascades in limited microscale or nanoscale environments has undergone a quick growth and attracted increasing interests in the field of rapid development of systems chemistry. In this study, alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH), and mesoporous silica nanoparticles (MSN) immobilized nicotinamide adenine dinucleotide (NAD+) were successfully immobilized on the zeolitic imidazolate frameworks (ZIFs). This immobilized product was named ZIF@ADH/NAD-MSN/LDH, and the effect of the multi-enzyme cascade was studied by measuring the catalytic synthesis of lactic acid. The loading efficiency of the enzyme in the in-situ co-immobilization method reached 92.65%. The synthesis rate of lactic acid was increased to 70.10%, which was about 2.82 times that of the free enzyme under the optimal conditions (40 °C, pH = 8). Additionally, ZIF@ADH/NAD-MSN/LDH had experimental stability (71.67% relative activity after four experiments) and storage stability (93.45% relative activity after three weeks of storage at 4 °C; 76.89% relative activity after incubation in acetonitrile-aqueous solution for 1 h; 27.42% relative activity after incubation in 15% N, N-Dimethylformamide (DMF) solution for 1 h). In summary, in this paper, the cyclic regeneration of coenzymes was achieved, and the reaction efficiency of the multi-enzyme biocatalytic cascade was improved due to the reduction of substrate diffusion.Keywords:confinement effect;multi-enzyme catalytic cascade;in-situ synthesis;coenzyme recycling;co-immobilization enzyme
研究领域:
生物化学
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纳米生物学
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系统化学
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