Simultaneous Engineering of the Thermostability and Activity of a Novel Aldehyde Dehydrogenase

ACS Catalysis, Год журнала: 2025, Номер 15(3), С. 1841 - 1853

Опубликована: Янв. 17, 2025

Acetaldehyde is a toxic pollutant that can be detoxified by acetaldehyde dehydrogenases (ADAs) through its conversion to acetyl-CoA. This study developed an integrated approach combining virtual screening, rational design, and dual scoring mechanism identify engineer hyperactive ADA variants. A library of 5000 Dickeya parazeae (DpADA) homologues was created protein BLAST, deep learning tools predicted their Kcat values. The top 100 candidates were selected based on binding affinity, evaluated molecular docking phylogenetic analysis. Among these, ADA6 from Buttiauxella sp. S04-F03 exhibited the highest activity, converting 57.6% acetyl-CoA, which 14.1 times higher than DpADA. To improve ADA6's thermostability, folding engineering applied, resulting in P443C variant with 80.7% increase residual activity after heat treatment. Molecular dynamics simulation pinpointed I440 as bottleneck substrate tunnel, guiding design dual-scoring system integrates structural adjustments electronic optimization evaluate mutations for improved exposure activity. final optimized variant, P443C-I440T, achieved efficiency 93.2%. demonstrates effectiveness computational mutagenesis enhance enzyme stability engineering.

Язык: Английский

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PROTACs: Novel tools for improving immunotherapy in cancer

Cancer Letters, Год журнала: 2023, Номер 560, С. 216128 - 216128

Опубликована: Март 16, 2023

Язык: Английский

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Drug discovery targeting nicotinamide phosphoribosyltransferase (NAMPT): Updated progress and perspectives

Bioorganic & Medicinal Chemistry, Год журнала: 2024, Номер 99, С. 117595 - 117595

Опубликована: Янв. 11, 2024

Язык: Английский

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Nicotinamide phosphoribosyltransferase prompts bleomycin-induced pulmonary fibrosis by driving macrophage M2 polarization in mice

Theranostics, Год журнала: 2024, Номер 14(7), С. 2794 - 2815

Опубликована: Янв. 1, 2024

Rationale: Idiopathic pulmonary fibrosis (IPF) is an irreversible, fatal interstitial lung disease lacking specific therapeutics.Nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting enzyme of nicotinamide adenine dinucleotide (NAD) salvage biosynthesis pathway and a cytokine, has been previously reported as biomarker for diseases; however, role NAMPT in not elucidated.Methods: We identified level changes by analyzing public RNA-Seq databases, verified collected clinical samples mice model Western blotting, qRT-PCR, ELISA Immunohistochemical staining.We investigated mechanism using pharmacological inhibition on Nampt transgenic mice.In vivo macrophage depletion clodronate liposomes reinfusion IL-4-induced M2 bone marrow-derived macrophages (BMDMs) from wild-type mice, combined with vitro cell experiments, were performed to further validate underlying involving fibrosis.Results: found that increased lungs patients IPF bleomycin (BLM)-induced fibrosis.NAMPT inhibitor FK866 alleviated BLM-induced significantly reduced levels bronchoalveolar lavage fluid (BALF).The single-cell RNA sequencing showed expression monocytes/macrophages was much higher than other cells.Knocking out mouse (Nampt fl/fl ;Cx3cr1 CreER ) decreased BALF, infiltration improved survival.Depleting subsequent BMDMs reversed protective effect monocyte/macrophage NAMPT-deletion fibrosis.In experiments confirmed engaged related released promoting polarization non-enzyme-dependent manner activating STAT6 signal pathway.Conclusions: prompts bleomycin-induced driving mice.Targeting promising strategy treating fibrosis.

Язык: Английский

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Targeting NAD + biosynthesis suppresses TGF-β1/Smads/RAB26 axis and potentiates cisplatin cytotoxicity in non-small cell lung cancer brain metastasis

Acta Neuropathologica Communications, Год журнала: 2025, Номер 13(1)

Опубликована: Март 11, 2025

Nicotinamide adenine dinucleotide (NAD+) plays an important role in tumor progression, but its non-small cell lung cancer with brain metastasis (NSCLC BM) remains unclear. Herein, we investigated NAD+ biosynthesis targeting as a new therapeutic strategy for NSCLC BM. Therapeutic activity of nicotinamide phosphoribosyl transferase (NAMPT) inhibitors was evaluated mouse models BM and using various assays such quantitation, viability, apoptosis assays. To explore impact on downstream signaling, RNA sequencing used NAMPT inhibitor-treated control cells, followed by validation genetic knockdown, western blot qRT-PCR. Expression proteins human association patient prognosis were examined. Finally, combination inhibitor cisplatin tested vivo. Systemic treatment demonstrated intracranial model. decreased cellular NAD levels suppressed proliferation invasion, induced cells. Supplementation precursor NMN rescued these effects. Mechanistically, disruption NAMPT-mediated TGF-β1/Smads/RAB26 leading to inhibition NAMPT/TGF-β1/Smads/RAB26 axis upregulated tissues correlated poor prognosis. Combining further extended the survival BM-bearing mice. Targeting provides can be effectively combined cisplatin. Our studies identified signaling NAMPT, which targeted mediate anti-cancer

Язык: Английский

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Innovative Cu/ZSM-5 nanostructures for development of a high-performance sensing platform for determination of NADH in biological fluids

Microchemical Journal, Год журнала: 2025, Номер unknown, С. 112972 - 112972

Опубликована: Фев. 1, 2025

Язык: Английский

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CE–MS Metabolomic and LC–MS Proteomic Analyses of Breast Cancer Exosomes Reveal Alterations in Purine and Carnitine Metabolism

Journal of Proteome Research, Год журнала: 2025, Номер unknown

Опубликована: Март 4, 2025

A nanosheath-flow capillary electrophoresis mass spectrometry (CE-MS) system with electrospray ionization was used to profile cationic metabolite cargo in exosomes secreted by nontumorigenic MCF-10A and tumorigenic MDA-MB-231 breast epithelial cells. An in-house-produced sheath liquid interface developed machined from PEEK enable nanoflow volumes. Normalization of CE-MS peak areas the total UV signal employed enhance quantitative accuracy reduce variability. CE-MS-based metabolomics revealed increased purine synthesis intermediates carnitine metabolites MDA-MB-231-derived exosomes, pathway enrichment indicating activation de novo pathways upregulation metabolism. In addition, nano-LC-MS-based proteomics differential expression ecto-5'-nucleotidase (NT5E) mitochondrial aldehyde dehydrogenase (ALDH9A1), demonstrating metabolic alterations related enzymatic steps. This study demonstrates application for comprehensive exosome metabolomics, uncovering reprogramming between normal cancerous cell lines providing insight into exosome-mediated signaling cancer

Язык: Английский

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Unveiling the future of cancer stem cell therapy: a narrative exploration of emerging innovations

Discover Oncology, Год журнала: 2025, Номер 16(1)

Опубликована: Март 22, 2025

Cancer stem cells (CSCs), are a critical subpopulation within tumours, and defined by their capacity for self-renewal, differentiation, tumour initiation. These unique traits contribute to progression, metastasis, resistance conventional treatments like chemotherapy radiotherapy, often resulting in cancer recurrence poor patient outcomes. As such, CSCs have become focal points developing advanced therapies. This review highlights progress CSC-targeted treatments, including chimeric antigen receptor T-cell (CAR-T) therapy, immunotherapy, molecular targeting, nanoparticle-based drug delivery systems. Plant-derived compounds gene-editing technologies, such as clustered regularly interspaced short palindromic repeats (CRISPR), explored potential enhance precision minimize side effects. Metabolic pathways integral CSC survival, mitochondrial dynamics, mitophagy (regulated dynamin-related protein 1 [DRP1] the PINK1/Parkin pathway), one-carbon metabolism, amino acid metabolism (involving enzymes glutaminase (GLS) glutamate dehydrogenase (GDH]), lipid hypoxia-induced metabolic reprogramming mediated hypoxia-inducible factors (HIF-1α HIF-2α), examined therapeutic targets. The adaptability of through autophagy, flexibility, epigenetic regulation metabolites α-ketoglutarate, succinate, fumarate is discussed. Additionally, extracellular vesicles nicotinamide adenine dinucleotide (NAD⁺) identified pivotal redox balance, DNA repair, modifications. Addressing challenges heterogeneity, immune evasion, treatment durability requires interdisciplinary collaboration. Advancing therapies essential overcoming preventing relapse, paving way transformative treatments. underscores importance leveraging innovative technologies fostering collaboration revolutionize treatment.

Язык: Английский

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The Role of P53 in Myocardial Ischemia-Reperfusion Injury

Cardiovascular Drugs and Therapy, Год журнала: 2023, Номер unknown

Опубликована: Июнь 30, 2023

Язык: Английский

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Use of ultra-high performance liquid chromatography-high-resolution mass spectroscopy to profile the metabolites from the serum of patients with breast cancer

Oncology Letters, Год журнала: 2024, Номер 27(5)

Опубликована: Март 14, 2024

Breast cancer (BC) is the most common type of malignancy and leading cause cancer-associated mortality in women worldwide. As such, assessing metabolic changes during human breast carcinogenesis key for developing disease prevention methods treatment. In present study, non-targeted metabolomics with chemometrics based on ultra-high performance liquid chromatography-high-resolution mass spectrometry were performed to assess differences serum metabolite patterns between patients BC healthy individuals. A total 3,246 metabolites sera controls found. Kyoto Encyclopedia Genes Genomes pathway analysis demonstrated that arginine, proline, nicotinate, nicotinamide, caffeine arachidonic acid metabolism, as well fatty biosynthesis significantly altered comparison controls. These results suggested profiling has potential discovering molecular biomarkers detection BC. It may also further understanding underlying mechanisms associated this disease.

Язык: Английский

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