Archives of biochemistry and biophysics最新文献

{"title":"Faecalibacterium prausnitzii alleviates Graves’ orbitopathy by modulating short-chain fatty acid (SCFA) metabolism and inhibiting orbital fibrosis and adipogenesis","authors":"Xin Qi ,&nbsp;Juzhong Zhang ,&nbsp;Yunping Li ,&nbsp;Yuxian Shi ,&nbsp;Jiaqi Yao ,&nbsp;Boding Tong","doi":"10.1016/j.abb.2025.110588","DOIUrl":"10.1016/j.abb.2025.110588","url":null,"abstract":"<div><div>Graves’ orbitopathy (GO) is an autoimmune inflammatory disorder characterized by orbital fibrosis and adipose tissue expansion, leading to proptosis and ocular dysfunction. Recent evidence suggests that gut microbiota dysbiosis contributes to GO pathogenesis, particularly through alterations in short-chain fatty acid (SCFA) metabolism. This study investigated the role of <em>Faecalibacterium prausnitzii</em> (<em>F. prausnitzii</em>), a key butyrate-producing bacterium, in modulating GO-related fibrosis and adipogenesis. Clinical fecal sample analysis revealed a significant reduction in <em>F. prausnitzii</em> abundance in GO patients. In a murine GO model, oral administration of <em>F. prausnitzii</em> significantly alleviated periorbital swelling, suppressed collagen deposition, and reduced lipid accumulation in orbital tissues. The metabolomic analysis demonstrated that <em>F. prausnitzii</em> restored butyrate levels, which were significantly depleted in GO. In vitro, butyrate treatment inhibited TGF-β-induced fibrosis and adipogenesis in orbital fibroblasts by downregulating α-SMA, collagen I, PPARγ, and C/EBPα expression. Additionally, <em>F. prausnitzii</em> administration improved thyroid function by reducing thyroid hormone and autoantibody levels. These findings highlight the therapeutic potential of <em>F. prausnitzii</em> in GO through modulation of gut microbiota-derived SCFA metabolism, suppression of fibroblast activation, and inhibition of adipogenic differentiation, providing a promising microbiota-based intervention for GO management.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110588"},"PeriodicalIF":3.0,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144854351","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Retraction notice to “Indirubin-3′-monoxime induces paraptosis in MDA-MB-231 breast cancer cells by transmitting Ca2+ from endoplasmic reticulum to mitochondria” [Arch. Biochem. Biophys. 698 (2021) 108723]","authors":"Matharage Gayani Dilshara ,&nbsp;Ilandarage Menu Neelaka Molagoda ,&nbsp;Rajapaksha Gedara Prasad Tharanga Jayasooriya ,&nbsp;Yung Hyun Choi ,&nbsp;Cheol Park ,&nbsp;Gi-Young Kim","doi":"10.1016/j.abb.2025.110578","DOIUrl":"10.1016/j.abb.2025.110578","url":null,"abstract":"","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110578"},"PeriodicalIF":3.0,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144844275","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Distinct interactions of proanthocyanidins of various degrees of polymerization with TLR4 receptor on the integrity of differentiating Caco-2 cell monolayers through tight junction protein expression","authors":"Wasitha P.D.W. Thilakarathna ,&nbsp;Madumani Amararathna ,&nbsp;Shakirah Azeez ,&nbsp;H.P. Vasantha Rupasinghe","doi":"10.1016/j.abb.2025.110592","DOIUrl":"10.1016/j.abb.2025.110592","url":null,"abstract":"<div><div>Many studies have demonstrated the potential of proanthocyanidins (PACs) to protect the intestinal epithelial barrier by regulating the cellular levels of tight junction (TJ) proteins. The effects of five PAC fractions with different mean degrees of polymerization (DP), namely, DT-PAC (dimers – trimers), OPAC-1 (oligomeric-PAC, pentamer – hexamer), OPAC-2 (octamer – dodecamer), PPAC-1 (polymeric-PAC, 13–15 subunits), and PPAC-2 (20 subunits), on the barrier integrity of differentiating Caco-2 cell monolayers were investigated. The effects of PAC fractions on the integrity of Caco-2 monolayers were evaluated by measuring the permeability of fluorescein isothiocyanate (FITC)-dextran and cellular levels of TJ proteins by western blotting. DT-PAC fraction favored the barrier integrity by significantly increasing the cellular protein levels of barrier-forming zonula occludens-1 (ZO-1) and claudin-3. The effects of OPAC fractions were controversial as the treatments simultaneously increased the cellular protein levels of barrier-forming claudin-1 and pore-forming claudin-2. The PPAC fractions significantly increased the permeation of FITC-dextran through Caco-2 monolayers, indicating a loss in barrier integrity. PPAC fractions disrupted the barrier integrity by reducing the cellular protein levels of barrier-forming ZO-1 and claudin-3 while increasing the pore-forming claudin-2. Molecular docking experiments revealed that PAC molecules with higher DP may disrupt Caco-2 monolayers by stimulating the cell surface toll-like receptor 4 (TLR4). Interestingly, PAC molecules with lower DP may inhibit the activation of TLR4 signaling. Highly polymerized PACs may disrupt the barrier integrity of intestinal epithelium undergoing differentiation by suppressing barrier-forming and upregulating the pore-forming TJ proteins, while less polymerized PACs protect barrier integrity by depicting opposing effects.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110592"},"PeriodicalIF":3.0,"publicationDate":"2025-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144858863","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"FSS in CTE triggers neuronal apoptosis through Piezo1-induced Ca2+ homeostasis disruption","authors":"Tianyu Gao ,&nbsp;Jingyuan Zhao ,&nbsp;Shuai Shao ,&nbsp;Yunong Yang ,&nbsp;Na Li ,&nbsp;Hong Yuan ,&nbsp;Bo Liu","doi":"10.1016/j.abb.2025.110587","DOIUrl":"10.1016/j.abb.2025.110587","url":null,"abstract":"<div><div>The altered cerebrospinal fluid dynamics induced by strenuous competitive sports result in repeated exposure of neurons to abnormal fluid shear stress (FSS). Although the stress intensity did not result in diffuse axonal injury, prolonged stimulation still induced chronic traumatic encephalopathy (CTE), characterized by neuronal dysfunction and apoptosis. However, the mechanism underlying the effects of elevated FSS in axon-intact cells remains unclear. In this study, microfluidic technology was utilized to apply FSS stimulation to depolarized SH-SY5Y cells. Utilizing Ca²⁺ biosensors based on Fluorescence resonance energy transfer technology to detect changes in intracellular calcium concentration ([Ca²⁺]_i) and endoplasmic reticulum Ca²⁺ release. The results indicated that FSS significantly elevate [Ca²⁺]_i during depolarization, enhancing vesicle release in short-term and apoptosis in long-term. The elevation of [Ca²⁺]_i was primarily attributed to extracellular Ca²⁺ influx via Piezo1 channels. Inhibition of Piezo1 activation suppressed aberrant vesicle release and attenuated apoptosis. This study identifies a novel CTE mechanism: FSS disrupts Ca²⁺ homeostasis in depolarized neurons via Piezo1-mediated Ca²⁺ influx, triggering aberrant vesicle release and apoptosis. This mechanism may provide crucial insights for the development of novel strategies to prevent or treat sports-related brain injuries and diseases.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110587"},"PeriodicalIF":3.0,"publicationDate":"2025-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144840707","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multiscale characterization of solar radiation-induced structural degradation in Type I collagen","authors":"Giuseppe De Luca ,&nbsp;Federica Piccirilli ,&nbsp;Olga Barrera ,&nbsp;Giuseppe Sancataldo ,&nbsp;Valeria Vetri","doi":"10.1016/j.abb.2025.110579","DOIUrl":"10.1016/j.abb.2025.110579","url":null,"abstract":"<div><div>Collagen, the most abundant structural protein in animals, plays a crucial role in maintaining skin integrity, elasticity, and strength. Type I collagen, which predominates in the skin, is particularly vulnerable to environmental stressors, such as solar radiation. Prolonged sun exposure accelerates collagen degradation, driving skin aging and impairing tissue functionality. However, the molecular mechanisms governing these intricate processes remain unclear. In this study, we employed bovine Type I collagen as a model system to investigate the molecular alterations induced by solar radiation, focusing on changes in structure, morphology, and fibrillogenesis potential. Collagen samples were irradiated using a solar simulator that mimics the full solar spectrum to ensure standardized conditions. Structural changes at different levels, were analyzed using a multi-technique approach combining classical spectroscopies, fluorescence lifetime imaging microscopy, and scattering-type scanning near-field optical microscopy (s-SNOM). This multimodal approach enabled both sensitive detection of molecular alterations and spatial mapping of local heterogeneities within collagen fibers. Results indicate partial destabilization of the triple-helical structure and a loss of cross-links and telopeptides, consistent with molecular misalignment. FLIM imaging on samples stained with Anilinonaphthalene-1-sulfonic acid (ANS), a gold standard fluorescent dye for the study of protein conformational transition highlighted increased sample heterogeneity and a reduction in hydrophobic regions, pointing to structural disruption which could be also related to the loss of self-assembly capabilities of collagen molecules.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110579"},"PeriodicalIF":3.0,"publicationDate":"2025-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144830228","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Higher confluency enhances lipid droplet size, triglyceride content and endocrine function on white-like adipocytes","authors":"Helen Yarimet Lorenzo-Anota ,&nbsp;Jorge E. Galván-Marroquín ,&nbsp;Antonio Sansén-Moreno ,&nbsp;Silvia López-Morán ,&nbsp;Jorge Benavides ,&nbsp;Karla Mayolo-Deloisa ,&nbsp;Marco Rito-Palomares ,&nbsp;Omar Lozano","doi":"10.1016/j.abb.2025.110575","DOIUrl":"10.1016/j.abb.2025.110575","url":null,"abstract":"<div><div>The murine fibroblast cell line 3T3-L1, due to their capability to differentiate into adipocytes, is extensively employed in studies of adipogenesis, metabolism and inflammation. However, the molecular phenotype and endocrine function is unclear with respect to initial confluency at the onset of the differentiation into adipocytes. This study aims to evaluate preadipocytes confluency implication during adipogenesis with morphological, biochemical, and molecular phenotype, and endocrine activity. Cells were stimulated by differentiation medium varying the initial confluency of either 75–85 % (low confluency group) or &gt;95 % (high confluency group) and were analyzed 10 and 14 days after the onset of the differentiation protocol, assessing lipid droplets, triglycerides, molecular phenotype, and endocrine function. The high confluency group showed higher frequencies of lipid droplets in the ranges 3.0–5.0 μm², 35–50 and &gt; 50 μm², with increased triglycerides synthesis at days 10 and 14, respectively, compared to either preadipocytes (2.51 ± 0.27 and 5.10 ± 2.09 -fold increase) or the low confluency group (2.51 ± 0.27 and 5.10 ± 2.09 -fold). Pref-1 expression decreased proportional to the differentiation time, and PGC-1α expression decreased on day 14 only for the high confluency group. PPARγ and adiponectin expression were directly proportional to confluency and time-differentiation. Adiponectin release increased independently of confluency and time, on the contrary, leptin release increased proportional to confluency and time-differentiation. In conclusion, a high confluency, compared to regular confluency, for white-like adipocyte differentiation increases lipid droplets biogenesis and triglycerides storage without altering molecular phenotype, enhancing endocrine function as observed by increased adiponectin and leptin release maintained up to 14 days. Our findings allow us to clarify the importance of confluency in the molecular phenotype and the endocrine function of white-like adipocytes.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"773 ","pages":"Article 110575"},"PeriodicalIF":3.0,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144783330","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Oncogenic RBM19 modulated by MAX-mediated allelic transcription regulation in hepatocellular carcinoma progression","authors":"Yanxiu Zheng ,&nbsp;Xiaolei Yin ,&nbsp;Linyu Han ,&nbsp;Shuqing Liu ,&nbsp;Nasha Zhang ,&nbsp;Yanting Yang ,&nbsp;Ming Yang","doi":"10.1016/j.abb.2025.110576","DOIUrl":"10.1016/j.abb.2025.110576","url":null,"abstract":"<div><div>Hepatocellular carcinoma (HCC), the most common type of liver cancer, exhibits the highest incidence in Asia and Africa. However, the mechanisms underlying the malignant transformation of normal cells to HCC cells remain poorly understood. MYC-associated protein X (MAX) functions as an oncogenic transcription factor (TF) in HCC by binding to E-box elements in promoters or enhancers of target gene. Using our previously developed Updated Integrative Functional Genomics Approach (TUIFGA), we identified 248 single nucleotide polymorphisms (SNPs) within MAX-binding sites in HCC cells. Among these SNPs, rs2290798 in the <em>RBM19</em> promoter was significantly associated with HBV-related HCC risk. Functional assays indicated that the rs2290798 genetic variant could disrupt TF MAX binding to the <em>RBM19</em> promoter and downregulate <em>RBM19</em> expression. RBM19 acts as an oncogenic RNA-binding protein (RBP) through promoting proliferation, migration, and invasion of HCC cells. Indeed, RBM19 functions as a RBP via stabilizing several oncogenic transcripts (<em>C1QTNF6</em>, <em>GLS1</em>, <em>MALAT1</em> or <em>RBM19</em>) and elevating their expression levels in HCC. These findings highlight the curial role of <em>RBM19</em> in HCC progression and its potential as a therapeutic target in cancers.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"772 ","pages":"Article 110576"},"PeriodicalIF":3.0,"publicationDate":"2025-08-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144780824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Correlation between methyltransferase METTL7B and atherosclerosis","authors":"Jian Xiong ,&nbsp;Xiaoyun Peng ,&nbsp;Liming Ma ,&nbsp;Fangcheng Zhu ,&nbsp;Yan Ding ,&nbsp;Zhixiao Wang","doi":"10.1016/j.abb.2025.110560","DOIUrl":"10.1016/j.abb.2025.110560","url":null,"abstract":"<div><div>Atherosclerosis (AS) is a serious threat to human health. Although glucose balance, lipid metabolism, inflammation and hypertension are closely related to AS, whether methyltransferase-like (METTL) family members are involved in the occurrence and development of AS remains elusive.</div><div>Differentially expressed genes of METTLs in AS and normal blood vessels in GSE43292 and GSE100927 databases were analyzed. Random forest screening was used to screen marker genes, and the intersection genes in the two databases were selected. GSE28829/GSE41571 and clinical tissue samples were used for verification. The databases were further used to analyze marker genes’ tissue and cellular localization and their correlation with lipid metabolism and efferocytosis.</div><div>7 and 17 differentially expressed METTL genes were obtained from GSE43292 and GSE100927 databases, respectively. METTL7B and METTL5 were verified as the intersection marker genes. Compared with the control group, the expression of METTL7B was significantly increased in advanced AS, AS ruptured plaque and clinical heavy-load plaque tissues. ROC curve analysis showed that the AUC of METTL7B in GSE28829 and GSE41571 was greater than 0.9. In addition, it was found that METTL7B was significantly correlated with lipid metabolism-related genes and promoted the formation of lipid droplets. METTL7B was positively correlated with atherosclerosis and macrophage-mediated efferocytosis. RNA-seq and targeted lipidomics results also confirmed that METTL7B is closely related to lipid metabolism and atherosclerosis. And further analysis also indicated that METTL7B could regulate 104 kinds of lipids, such as Lipid-n-0041, Lipid-n-0056, Lipid-n-0057, Lipid-n-0098, Lipid-n-0099 and Lipid-n-0169, mediated by AKR1C1, CETP and RORA. This study reveals a new mechanism for the occurrence and development of AS, thereby providing a potential target for the treatment of AS.</div><div>In conclusion, METTL7B can be used as a predictor and therapeutic target for AS.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"772 ","pages":"Article 110560"},"PeriodicalIF":3.0,"publicationDate":"2025-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144757355","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dexmedetomidine attenuates oxaliplatin-induced neuropathic pain by modulating the TLR4/NF-κB pathway to reduce spinal inflammation and oxidative stress","authors":"Yuchao Lin,&nbsp;Kexin Chen,&nbsp;Lei Zhao,&nbsp;Ming Zhao,&nbsp;Yuanyuan Liu,&nbsp;Yu Li","doi":"10.1016/j.abb.2025.110572","DOIUrl":"10.1016/j.abb.2025.110572","url":null,"abstract":"<div><h3>Purpose</h3><div>This study aimed to investigate the effects of Dexmedetomidine (Dex) on oxaliplatin-induced neuropathic pain and its underlying mechanisms.</div></div><div><h3>Methods</h3><div>A murine model of oxaliplatin-induced neuropathic pain was established using intraperitoneal injections of oxaliplatin. Dex was administered at different doses, and behavioral tests were performed to assess pain. Spinal cord tissues were analyzed for inflammatory cytokines, oxidative stress markers, and key signaling molecules related to the toll-like receptor 4 (TLR4)/nuclear factor kappa B using quantitative real-time polymerase chain reaction (qRT-PCR), Western blot, and immunohistochemistry. In addition, in vitro experiments were conducted using TNF-α-stimulated C6 glial cells to further assess the anti-inflammatory effects of Dex.</div></div><div><h3>Results</h3><div>Dex significantly alleviated oxaliplatin-induced neuropathic pain, as shown by an increase in paw withdrawal thresholds and a marked reduction in spontaneous flinching. Molecular analyses further demonstrated that Dex treatment reduced interleukin-1 beta (IL-1β), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) expression, as well as malondialdehyde (MDA) and cyclooxygenase-2 (COX2) in the spinal cord. Concurrently, there was a notable enhancement in the activity of Manganese superoxide dismutase (Mn-SOD) and glutathione (GSH), suggesting improved antioxidative defense. Additionally, Dex reduced spinal inflammation and oxidative stress by downregulating TLR4 expression and inhibiting NF-κB activation. Consistent with these findings, Dex also suppressed NF-κB phosphorylation and cytokine expression in TNF-α-treated C6 cells in vitro.</div></div><div><h3>Conclusions</h3><div>Dex significantly reduced oxaliplatin-induced neuropathic pain by downregulating TLR4 expression and inhibiting NF-κB activation.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"772 ","pages":"Article 110572"},"PeriodicalIF":3.0,"publicationDate":"2025-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144759000","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular insights into CYP19A1 mutations and their role in estrogen production","authors":"Yoo-bin Lee,&nbsp;Changmin Kim,&nbsp;Jiyeon Hong,&nbsp;Donghak Kim","doi":"10.1016/j.abb.2025.110573","DOIUrl":"10.1016/j.abb.2025.110573","url":null,"abstract":"<div><div>Human cytochrome P450 enzyme CYP19A1, commonly referred to as aromatase, plays a critical role in estrogen biosynthesis by catalyzing the conversion of C19 androgens into aromatic C18 estrogens. Alterations in aromatase activity have been implicated in the development and progression of estrogen-dependent diseases. Genetic mutations, including nonsynonymous single nucleotide polymorphisms (SNPs), can markedly impact the catalytic function of CYP19A1. In this study, we investigated the functional implications of six CYP19A1 variants (R192H, R192Q, T201M, R264C, P308F, and M364T). These variants were generated via site-directed mutagenesis, expressed in <em>Escherichia coli</em>, and purified. The T201M, R264C, and P308F variants exhibited measurable expression levels ranging from 90 to 150nmol P450 per liter culture, whereas no detectable P450 holoenzyme was observed for the R192H, R192Q, and M364T variants. Spectral binding assays revealed typical type I spectral shifts upon androgen binding for all purified variants with tight affinity. Catalytic activities of testosterone and androstenedione aromatization were assessed by UPLC–mass spectrometry, and steady-state kinetic analyses demonstrated that T201M and R264C retained catalytic efficiencies comparable to wild-type, with efficiency ratios of 1.1–0.9 for androstenedione and 0.6–0.8 for testosterone, respectively, relative to the wild-type. In contrast, P308F showed marked reductions in catalytic efficiencies (0.2 and 0.1), driven by decreases in both <em>k</em>_cat and <em>K</em>_m values. Structural analysis indicates that the proline residue at position 308, situated within the I-helix, is likely critical for stabilizing a catalytically active conformation of the enzyme-substrate complex. These results provide valuable insights into the functional role of CYP19A1 in estrogen biosynthesis and could inform the design of innovative therapies for estrogen-related diseases.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"772 ","pages":"Article 110573"},"PeriodicalIF":3.0,"publicationDate":"2025-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144723950","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}