C-X-C motif chemokine ligand 17 (CXCL17) is a chemoattractant whose receptor remains controversial. While recent studies identified CXCL17 as an agonist of G protein-coupled receptor 25 (GPR25), it was also reported to activate MAS-related GPR family member X2 (MRGPRX2), a member of MAS-related G protein-coupled receptors (MRGPRs), though this finding has not yet been reproduced by other laboratories. In this study, we confirmed that micromolar concentrations of human CXCL17 activate human MRGPRX2 in transfected human embryonic kidney (HEK) 293T cells using a NanoLuc Binary Technology (NanoBiT)-based β-arrestin recruitment assay. We further demonstrated that human CXCL17 also activates MRGPRX1 and MAS1 among 10 human MRGPRs in the same assay. CXCL17 could also induce chemotactic movement of transfected HEK293T cells expressing MRGPRX2, MRGPRX1, or MAS1. However, removal of C-terminal residues from CXCL17 did not affect its activation of these three MRGPRs, even though this region is essential for GPR25 activation. These results suggest that CXCL17 activates MRGPRX2, MRGPRX1, and MAS1 through a mechanism distinct from GPR25 activation. Further investigation is needed to determine whether these MRGPRs mediate the in vivo functions of CXCL17.
{"title":"CXCL17 activates three MAS-related G protein-coupled receptors independently of its conserved C-terminal fragment","authors":"Wen-Feng Hu, Juan-Juan Wang, Jie Yu, Ya-Li Liu, Zeng-Guang Xu, Zhan-Yun Guo","doi":"10.1016/j.abb.2025.110666","DOIUrl":"10.1016/j.abb.2025.110666","url":null,"abstract":"<div><div>C-X-C motif chemokine ligand 17 (CXCL17) is a chemoattractant whose receptor remains controversial. While recent studies identified CXCL17 as an agonist of G protein-coupled receptor 25 (GPR25), it was also reported to activate MAS-related GPR family member X2 (MRGPRX2), a member of MAS-related G protein-coupled receptors (MRGPRs), though this finding has not yet been reproduced by other laboratories. In this study, we confirmed that micromolar concentrations of human CXCL17 activate human MRGPRX2 in transfected human embryonic kidney (HEK) 293T cells using a NanoLuc Binary Technology (NanoBiT)-based β-arrestin recruitment assay. We further demonstrated that human CXCL17 also activates MRGPRX1 and MAS1 among 10 human MRGPRs in the same assay. CXCL17 could also induce chemotactic movement of transfected HEK293T cells expressing MRGPRX2, MRGPRX1, or MAS1. However, removal of C-terminal residues from CXCL17 did not affect its activation of these three MRGPRs, even though this region is essential for GPR25 activation. These results suggest that CXCL17 activates MRGPRX2, MRGPRX1, and MAS1 through a mechanism distinct from GPR25 activation. Further investigation is needed to determine whether these MRGPRs mediate the <em>in vivo</em> functions of CXCL17.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"775 ","pages":"Article 110666"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145407900","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}
Pub Date : 2026-01-01Epub Date: 2025-10-27DOI: 10.1016/j.abb.2025.110665
Mutsumi Yoshida , Yuya Hanazono , Nobutaka Numoto , Saaya Yabuno , Nobutoshi Ito , Takachika Azuma , Masayuki Oda
The phenomenon in which the antibody affinity for T cell-dependent antigens increases through multiple rounds of somatic hypermutation (SHM) is referred to as affinity maturation. The elucidation of the structural and physical properties of antibodies obtained at various stages of the affinity maturation process can help us understand the molecular recognition mechanism of proteins in general. For this purpose, we used anti-(4-hydroxy-3-nitrophenyl)acetyl (NP) single-chain (scFv) antibodies, prepared from the parent antibodies F8, B2, C6, and E11, and analyzed the crystal structures either in the absence or presence of NP or (4-hydroxy-3,5-dinitrophenyl)acetyl (NNP). Comparison of the structures revealed that the antibodies shared a common antigen recognition architecture consisting of residues with basic side chains, Arg50H and Lys58H/Arg58H, in addition to those at the junctional positions of gene segments, Trp96L and Tyr95H or His100BH. These residues are responsible for the recognition of antigenic determinants, nitro-, hydroxyl- and phenylacetyl-groups, through hydrogen bond formation. Second, the Trp33H to Leu33H mutation seemed to strengthen the interaction because the antigen was closer to the combining site. Finally, analysis of NP and NNP complexes showed little difference in the antigen recognition modes and in the overall structures of the complementarity-determining regions between C6 and E11 scFvs. It was suggested that the replacement of residues by SHM provided a unique binding site for each antibody by fine tuning the microenvironment without disturbing specificity.
{"title":"Structural basis of affinity maturation of anti-(4-hydroxy-3-nitrophenyl)acetyl antibodies","authors":"Mutsumi Yoshida , Yuya Hanazono , Nobutaka Numoto , Saaya Yabuno , Nobutoshi Ito , Takachika Azuma , Masayuki Oda","doi":"10.1016/j.abb.2025.110665","DOIUrl":"10.1016/j.abb.2025.110665","url":null,"abstract":"<div><div>The phenomenon in which the antibody affinity for T cell-dependent antigens increases through multiple rounds of somatic hypermutation (SHM) is referred to as affinity maturation. The elucidation of the structural and physical properties of antibodies obtained at various stages of the affinity maturation process can help us understand the molecular recognition mechanism of proteins in general. For this purpose, we used anti-(4-hydroxy-3-nitrophenyl)acetyl (NP) single-chain (scFv) antibodies, prepared from the parent antibodies F8, B2, C6, and E11, and analyzed the crystal structures either in the absence or presence of NP or (4-hydroxy-3,5-dinitrophenyl)acetyl (NNP). Comparison of the structures revealed that the antibodies shared a common antigen recognition architecture consisting of residues with basic side chains, Arg50<sup>H</sup> and Lys58<sup>H</sup>/Arg58<sup>H</sup>, in addition to those at the junctional positions of gene segments, Trp96<sup>L</sup> and Tyr95<sup>H</sup> or His100B<sup>H</sup>. These residues are responsible for the recognition of antigenic determinants, nitro-, hydroxyl- and phenylacetyl-groups, through hydrogen bond formation. Second, the Trp33<sup>H</sup> to Leu33<sup>H</sup> mutation seemed to strengthen the interaction because the antigen was closer to the combining site. Finally, analysis of NP and NNP complexes showed little difference in the antigen recognition modes and in the overall structures of the complementarity-determining regions between C6 and E11 scFvs. It was suggested that the replacement of residues by SHM provided a unique binding site for each antibody by fine tuning the microenvironment without disturbing specificity.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"775 ","pages":"Article 110665"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145399720","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}
Pub Date : 2026-01-01Epub Date: 2025-10-24DOI: 10.1016/j.abb.2025.110650
Elena G. Varlamova , Vladimir V. Rogachev , Sergey V. Gudkov , Egor A. Turovsky
Background
The neuroprotective potential of tellurium nanoparticles (TeNPs) remains critically underexplored compared to their selenium analogues, despite their presumed physicochemical similarities. A significant limitation has been the absence of a reliable method for producing highly stable, monodisperse TeNPs without chemical contaminants for robust biological assessment.
Key results
In this study, we developed a novel laser ablation protocol to synthesize stable spherical TeNPs with two distinct diameters (10 nm and 100 nm). We comprehensively investigated their diameter-dependent effects on primary cortical cells under excitotoxicity and ischemia models. Our key finding reveals a striking dichotomy of effects: 100 nm-sized TeNPs conferred significant cytoprotection against glutamate excitotoxicity, primarily by suppressing necrotic cell death. In contrast, 10 nm-sized TeNPs exacerbated oxidative stress, upregulated pro-apoptotic and pro-inflammatory gene expression, and failed to provide protection in either injury model. Furthermore, 100 nm-sized TeNPs uniquely modulated selenoprotein gene expression patterns under ischemic stress. Surprisingly, particles of neither diameter protected against oxygen-glucose deprivation (OGD), while 100 nm-sized TeNPs potentiated ischemia-induced calcium overload.
Originality and significance
This work is the first to (1) employ laser-ablated TeNPs in neurobiological research, (2) demonstrate a complete reversal of neuroprotective outcomes based solely on nanoparticle size, and (3) establish correlations between TeNP-induced effects and specific alterations in calcium signaling, reactive oxygen species (ROS) production, and gene expression profiles in brain cells. Our findings underscore that nanoparticle size is a critical, non-negotiable parameter determining the therapeutic-to-toxic outcome of TeNPs in the brain. This provides a crucial foundation for the future design of nanotherapeutic agents for neurological disorders.
{"title":"From toxicity to protection: Size-specific effects of tellurium nanoparticles on cortical cell survival","authors":"Elena G. Varlamova , Vladimir V. Rogachev , Sergey V. Gudkov , Egor A. Turovsky","doi":"10.1016/j.abb.2025.110650","DOIUrl":"10.1016/j.abb.2025.110650","url":null,"abstract":"<div><h3>Background</h3><div>The neuroprotective potential of tellurium nanoparticles (TeNPs) remains critically underexplored compared to their selenium analogues, despite their presumed physicochemical similarities. A significant limitation has been the absence of a reliable method for producing highly stable, monodisperse TeNPs without chemical contaminants for robust biological assessment.</div></div><div><h3>Key results</h3><div>In this study, we developed a novel laser ablation protocol to synthesize stable spherical TeNPs with two distinct diameters (10 nm and 100 nm). We comprehensively investigated their diameter-dependent effects on primary cortical cells under excitotoxicity and ischemia models. Our key finding reveals a striking dichotomy of effects: 100 nm-sized TeNPs conferred significant cytoprotection against glutamate excitotoxicity, primarily by suppressing necrotic cell death. In contrast, 10 nm-sized TeNPs exacerbated oxidative stress, upregulated pro-apoptotic and pro-inflammatory gene expression, and failed to provide protection in either injury model. Furthermore, 100 nm-sized TeNPs uniquely modulated selenoprotein gene expression patterns under ischemic stress. Surprisingly, particles of neither diameter protected against oxygen-glucose deprivation (OGD), while 100 nm-sized TeNPs potentiated ischemia-induced calcium overload.</div></div><div><h3>Originality and significance</h3><div>This work is the first to (1) employ laser-ablated TeNPs in neurobiological research, (2) demonstrate a complete reversal of neuroprotective outcomes based solely on nanoparticle size, and (3) establish correlations between TeNP-induced effects and specific alterations in calcium signaling, reactive oxygen species (ROS) production, and gene expression profiles in brain cells. Our findings underscore that nanoparticle size is a critical, non-negotiable parameter determining the therapeutic-to-toxic outcome of TeNPs in the brain. This provides a crucial foundation for the future design of nanotherapeutic agents for neurological disorders.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"775 ","pages":"Article 110650"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145414502","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}
Pub Date : 2026-01-01Epub Date: 2025-11-24DOI: 10.1016/j.abb.2025.110677
Duygu İnci Özbağcı , Sevinç İlkar Erdağı , Rahmiye Aydın
Aromatic secondary metabolites, hydroxybenzoic acids (HBAs)—β-resorcylic (2,4-dihydroxybenzoic acid) and gallic acid (3,4,5-trihydroxybenzoic acid)—constitute a class of phenolic compounds with numerous bioactivities that are of pharmacological and therapeutic interest. Urease plays a critical role in various biological pathways, with its activation linked to medical conditions including nephrolithiasis and peptic ulcers. The pursuit of potent and safe urease inhibitors has become a key priority in the field of pharmaceutical research. The present study sought to explore the molecular interactions between HBAs and urease to better understand their binding behavior. To assess the impact of both HBAs on the structural integrity and enzymatic activity of urease, fluorescence spectroscopy, FTIR, and molecular docking were utilized. Both HBAs quenched urease fluorescence through static mechanisms and bound favorably to the Ni2+-centered catalytic site. In silico ADME and toxicity profiling confirmed their drug-likeness and low predicted toxicity. Gallic acid exhibited stronger antioxidant and urease inhibition activity than β-resorcylic acid, highlighting the importance of hydroxyl substituents in radical scavenging. These findings suggest HBAs as promising natural scaffolds for developing dual-function urease inhibitors with antioxidant potential.
{"title":"Aromatic secondary metabolite interactions with nickel-dependent urease","authors":"Duygu İnci Özbağcı , Sevinç İlkar Erdağı , Rahmiye Aydın","doi":"10.1016/j.abb.2025.110677","DOIUrl":"10.1016/j.abb.2025.110677","url":null,"abstract":"<div><div>Aromatic secondary metabolites, hydroxybenzoic acids (HBAs)—β-resorcylic (2,4-dihydroxybenzoic acid) and gallic acid (3,4,5-trihydroxybenzoic acid)—constitute a class of phenolic compounds with numerous bioactivities that are of pharmacological and therapeutic interest. Urease plays a critical role in various biological pathways, with its activation linked to medical conditions including nephrolithiasis and peptic ulcers. The pursuit of potent and safe urease inhibitors has become a key priority in the field of pharmaceutical research. The present study sought to explore the molecular interactions between HBAs and urease to better understand their binding behavior. To assess the impact of both HBAs on the structural integrity and enzymatic activity of urease, fluorescence spectroscopy, FTIR, and molecular docking were utilized. Both HBAs quenched urease fluorescence through static mechanisms and bound favorably to the Ni<sup>2+</sup>-centered catalytic site. In silico ADME and toxicity profiling confirmed their drug-likeness and low predicted toxicity. Gallic acid exhibited stronger antioxidant and urease inhibition activity than β-resorcylic acid, highlighting the importance of hydroxyl substituents in radical scavenging. These findings suggest HBAs as promising natural scaffolds for developing dual-function urease inhibitors with antioxidant potential.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"775 ","pages":"Article 110677"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145621155","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}
Pub Date : 2026-01-01Epub Date: 2025-10-25DOI: 10.1016/j.abb.2025.110653
Zhaohui Yang , Jiawen Hao , Zhongyu Ma , Yunxiao Xie , Ruixue Zhang , Xuewei Yin , Guimin Wang , Yinqiao Zhang , Miao Zhang , Xuan Zhang , Dadong Guo , Hongsheng Bi
Objective
To explore the influence of the activated Notch1 signaling pathway on choroidal tissue fibrosis in experimental myopia.
Methods
Right eyes were fitted with a −6.0D lens to create experimental myopia. Ocular parameters, choroidal thickness, and blood flow were assessed after the injection of shRNA-Notch1 and the Notch1 pathway inhibitor DAPT into the vitreous cavity. Changes in blood flow in the choroid were observed using optical coherence tomography angiography (OCTA). The expression levels of molecules associated with the Notch1 signaling pathway were assessed at both the gene and protein levels. Additionally, we examined the choroidal thickness, level of choroidal fibrosis, and level of apoptosis using hematoxylin and eosin (H&E) staining, Masson staining, and TdT-mediated dUTP Nick-End Labeling (TUNEL).
Results
Compared with the NC group, the LIM group presented a substantial decrease in diopter, increased optical axis length, decreased choroidal thickness, and significantly reduced blood perfusion. qPCR and Western blotting revealed that the expression levels of Notch1, HES1, DLL1, TGF-β, α-SMA, and collagen Ⅰ in the LIM group were considerably greater than those in the NC group. Masson staining confirmed that fibrosis occurred in the choroidal tissue of the LIM group. Vitreous injection of shRNA-Notch1 and DAPT attenuated fibrosis, thereby controlling myopia progression.
Conclusion
Activation of the Notch1 signaling pathway leads to choroidal physiological dysfunction and elongated optical axis length, driving the progression of myopia. By inhibiting the Notch1 signaling pathway, shRNA-Notch1 can suppress optical axis elongation, increase choroidal thickness, and attenuate choroidal fibrosis, thus inhibiting the progression of myopia.
{"title":"Aggravated choroidal tissue fibrosis in experimental myopia is involved in activation of the Notch1 signaling pathway","authors":"Zhaohui Yang , Jiawen Hao , Zhongyu Ma , Yunxiao Xie , Ruixue Zhang , Xuewei Yin , Guimin Wang , Yinqiao Zhang , Miao Zhang , Xuan Zhang , Dadong Guo , Hongsheng Bi","doi":"10.1016/j.abb.2025.110653","DOIUrl":"10.1016/j.abb.2025.110653","url":null,"abstract":"<div><h3>Objective</h3><div>To explore the influence of the activated Notch1 signaling pathway on choroidal tissue fibrosis in experimental myopia.</div></div><div><h3>Methods</h3><div>Right eyes were fitted with a −6.0D lens to create experimental myopia. Ocular parameters, choroidal thickness, and blood flow were assessed after the injection of shRNA-Notch1 and the Notch1 pathway inhibitor DAPT into the vitreous cavity. Changes in blood flow in the choroid were observed using optical coherence tomography angiography (OCTA). The expression levels of molecules associated with the Notch1 signaling pathway were assessed at both the gene and protein levels. Additionally, we examined the choroidal thickness, level of choroidal fibrosis, and level of apoptosis using hematoxylin and eosin (H&E) staining, Masson staining, and TdT-mediated dUTP Nick-End Labeling (TUNEL).</div></div><div><h3>Results</h3><div>Compared with the NC group, the LIM group presented a substantial decrease in diopter, increased optical axis length, decreased choroidal thickness, and significantly reduced blood perfusion. qPCR and Western blotting revealed that the expression levels of Notch1, HES1, DLL1, TGF-β, α-SMA, and collagen Ⅰ in the LIM group were considerably greater than those in the NC group. Masson staining confirmed that fibrosis occurred in the choroidal tissue of the LIM group. Vitreous injection of shRNA-Notch1 and DAPT attenuated fibrosis, thereby controlling myopia progression.</div></div><div><h3>Conclusion</h3><div>Activation of the Notch1 signaling pathway leads to choroidal physiological dysfunction and elongated optical axis length, driving the progression of myopia. By inhibiting the Notch1 signaling pathway, shRNA-Notch1 can suppress optical axis elongation, increase choroidal thickness, and attenuate choroidal fibrosis, thus inhibiting the progression of myopia.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"775 ","pages":"Article 110653"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145414929","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}
Pub Date : 2026-01-01Epub Date: 2025-10-21DOI: 10.1016/j.abb.2025.110642
Maria G. Rivas , Raquel S. Correia Cordeiro , Rashmi R. Nair , Lindomar J.C. Albuquerque , Márcia A.S. Correia , Alejandro Panjkovich , Dmitri I. Svergun , Teresa Santos-Silva
Molybdenum and tungsten are taken up by cells through highly specific transport systems known in bacteria as ModABC and Tup/WtpABC, respectively. Component A (Mod/Tup/WtpA) binds the metal in the periplasm and thus represents the first selective mechanism for the uptake of the correct metal. The genome of Oleidesulfovibrio alaskensis G20 contains both mod and tup genes. In a previous paper, we reported the structure, molybdenum and tungsten-binding constants, and biochemical properties of TupA isolated from this organism. In the current study, we used biochemical and spectroscopic techniques to explore changes in the structure and stability of ModA upon metal binding. We show that the OaModA is expressed as a monomeric and soluble protein. In addition, it binds both molybdate and tungstate with extremely high affinity with KD constant in the nanomolar range and is unable to distinguish between them. The protein is more stable upon molybdenum binding, as demonstrated by differential scanning fluorescence studies. Such change is accompanied by a transition to a closed conformation with changes in secondary structure.
{"title":"Studies of the structural properties and stability of the molybdenum transport protein ModA from Oleidesulfovibrio alaskensis G20 upon metal binding","authors":"Maria G. Rivas , Raquel S. Correia Cordeiro , Rashmi R. Nair , Lindomar J.C. Albuquerque , Márcia A.S. Correia , Alejandro Panjkovich , Dmitri I. Svergun , Teresa Santos-Silva","doi":"10.1016/j.abb.2025.110642","DOIUrl":"10.1016/j.abb.2025.110642","url":null,"abstract":"<div><div>Molybdenum and tungsten are taken up by cells through highly specific transport systems known in bacteria as ModABC and Tup/WtpABC, respectively. Component A (Mod/Tup/WtpA) binds the metal in the periplasm and thus represents the first selective mechanism for the uptake of the correct metal. The genome of <em>Oleidesulfovibrio alaskensis</em> G20 contains both mod and tup <em>genes</em>. In a previous paper, we reported the structure, molybdenum and tungsten-binding constants, and biochemical properties of TupA isolated from this organism. In the current study, we used biochemical and spectroscopic techniques to explore changes in the structure and stability of ModA upon metal binding. We show that the <em>Oa</em>ModA is expressed as a monomeric and soluble protein. In addition, it binds both molybdate and tungstate with extremely high affinity with <em>K</em><sub>D</sub> constant in the nanomolar range and is unable to distinguish between them. The protein is more stable upon molybdenum binding, as demonstrated by differential scanning fluorescence studies. Such change is accompanied by a transition to a closed conformation with changes in secondary structure.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"775 ","pages":"Article 110642"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145353451","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}
Pub Date : 2026-01-01Epub Date: 2025-10-29DOI: 10.1016/j.abb.2025.110651
Qun Gao , Qi Wang , Yanjiao Hu , Fangjie Xin , Jie Chen , Sha Yu , Xinli Chen , Qingqing Lv , Baoxia Cui , Jun Jiao , Xinlin Jiao
Purpose
To investigate the impact of SIRT7 on the development of cervical cancer and its relationship with cuproptosis in cervical cancer.
Method
HeLa and SiHa cells were transfected with lentiviruses for SIRT7 overexpression and knockdown. The effects of SIRT7 on cervical cancer cell proliferation, apoptosis, invasion, and migration were analyzed using CCK8, plate cloning, flow cytometry, Transwell assays, and scratch assays. To verify the relationship between SIRT7 and cuproptosis, we utilized cuproptosis inhibitors and activators. Immunofluorescence, transmission electron microscope, flow cytometry, ELISA, and Western blot were used to analyze copper ion content, mitochondrial ultrastructure, cellular reactive oxygen species, mitochondrial membrane potential, pyruvate levels, cell viability and the cuproptosis-related proteins.
Results
SIRT7 enhanced the proliferation, migration, and invasion of HeLa and SiHa cells, inhibited apoptosis, and promoted cervical cancer growth. Knocking down SIRT7 caused cuproptosis of HeLa and SiHa cells, characterized by increased Cu2+ content, disrupted mitochondrial structure, decreased membrane potential, elevated ROS production, and upregulation of cuproptosis-related proteins SLC31A1 and HSP70, and downregulation of FDX1, LIAS and DLAT. Low SIRT7 expression's effect on cuproptosis was reduced by TTM. SIRT7 overexpression inhibited cuproptosis, unlike SIRT7 knockdown. SIRT7 overexpression's inhibitory effect on cuproptosis is altered by rhSLC31A1.
Conclusion
SIRT7 was recognized as an oncogene in cervical cancer, which boosted cervical cancer cell proliferation and invasion, lowered intracellular copper levels, and prevented cuproptosis. SIRT7 downregulation triggered cuproptosis, inhibiting tumor cell growth.
{"title":"SIRT7 deficiency promoted cuproptosis-mediated mitochondrial dysfunction and inhibited malignant development of cervical cancer","authors":"Qun Gao , Qi Wang , Yanjiao Hu , Fangjie Xin , Jie Chen , Sha Yu , Xinli Chen , Qingqing Lv , Baoxia Cui , Jun Jiao , Xinlin Jiao","doi":"10.1016/j.abb.2025.110651","DOIUrl":"10.1016/j.abb.2025.110651","url":null,"abstract":"<div><h3>Purpose</h3><div>To investigate the impact of SIRT7 on the development of cervical cancer and its relationship with cuproptosis in cervical cancer.</div></div><div><h3>Method</h3><div>HeLa and SiHa cells were transfected with lentiviruses for SIRT7 overexpression and knockdown. The effects of SIRT7 on cervical cancer cell proliferation, apoptosis, invasion, and migration were analyzed using CCK8, plate cloning, flow cytometry, Transwell assays, and scratch assays. To verify the relationship between SIRT7 and cuproptosis, we utilized cuproptosis inhibitors and activators. Immunofluorescence, transmission electron microscope, flow cytometry, ELISA, and Western blot were used to analyze copper ion content, mitochondrial ultrastructure, cellular reactive oxygen species, mitochondrial membrane potential, pyruvate levels, cell viability and the cuproptosis-related proteins.</div></div><div><h3>Results</h3><div>SIRT7 enhanced the proliferation, migration, and invasion of HeLa and SiHa cells, inhibited apoptosis, and promoted cervical cancer growth. Knocking down SIRT7 caused cuproptosis of HeLa and SiHa cells, characterized by increased Cu<sup>2+</sup> content, disrupted mitochondrial structure, decreased membrane potential, elevated ROS production, and upregulation of cuproptosis-related proteins SLC31A1 and HSP70, and downregulation of FDX1, LIAS and DLAT. Low SIRT7 expression's effect on cuproptosis was reduced by TTM. SIRT7 overexpression inhibited cuproptosis, unlike SIRT7 knockdown. SIRT7 overexpression's inhibitory effect on cuproptosis is altered by rhSLC31A1.</div></div><div><h3>Conclusion</h3><div>SIRT7 was recognized as an oncogene in cervical cancer, which boosted cervical cancer cell proliferation and invasion, lowered intracellular copper levels, and prevented cuproptosis. SIRT7 downregulation triggered cuproptosis, inhibiting tumor cell growth.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"775 ","pages":"Article 110651"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145407881","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}
Pub Date : 2026-01-01Epub Date: 2025-10-28DOI: 10.1016/j.abb.2025.110667
Sheng Lin, Junbin Chen
Background
Dysregulated microRNAs are implicated in the acute cerebral infarction (ACI) pathogenesis.
Aim
This study investigated the clinical significance and mechanisms of miR-943 in ACI.
Methods
Serum miR-943 levels were measured using RT-qPCR in 132 ACI patients and 135 healthy controls (HC). Diagnostic potential was evaluated by ROC analysis. Prognosis was assessed using 90-day mRS scores. Correlations with clinical parameters were analyzed using Pearson's/Spearman's tests. Multivariate logistic regression identified outcome predictors. Bioinformatic prediction and experimental validation (dual-luciferase, RIP assays) identified miR-943 targets. An OGD/R model in BV-2 microglia simulated ischemia-reperfusion. Effects of miR-943 inhibitors and si-BDNF on viability (CCK-8), apoptosis (flow cytometry), and inflammation (ELISA) were assessed.
Results
ACI patients exhibited significantly elevated serum miR-943 expression and reduced BDNF levels versus HC. miR-943 showed high diagnostic value (AUC = 0.909, sensitivity 85.6 %, specificity 83.7 %). MiR-943 expression correlated positively with SBP, DBP, FBG, Hcy, NIHSS score, and infarct volume. Patients with a poor prognosis exhibited significantly higher miR-943 expression levels. Multivariate analysis identified high expression of miR-943 as an independent risk factor for poor outcome. BDNF was validated as a direct miR-943 target gene. OGD/R increased miR-943 expression, suppressed proliferation, promoted apoptosis, and enhanced the expression of inflammatory factors in BV-2 cells. Inhibiting miR-943 expression reversed these detrimental effects, enhancing proliferation and reducing apoptosis/inflammation. Critically, BDNF knockdown abolished the protective effects of miR-943 inhibition.
Conclusion
Serum miR-943 is a promising diagnostic and prognostic biomarker for ACI. It exerts pathological effects by post-transcriptionally suppressing BDNF. Targeting the miR-943/BDNF axis represents a potential therapeutic strategy.
{"title":"Clinical value of serum miR-943 as a diagnostic and prognostic biomarker for acute cerebral infarction and its mechanism of action via targeting BDNF","authors":"Sheng Lin, Junbin Chen","doi":"10.1016/j.abb.2025.110667","DOIUrl":"10.1016/j.abb.2025.110667","url":null,"abstract":"<div><h3>Background</h3><div>Dysregulated microRNAs are implicated in the acute cerebral infarction (ACI) pathogenesis.</div></div><div><h3>Aim</h3><div>This study investigated the clinical significance and mechanisms of miR-943 in ACI.</div></div><div><h3>Methods</h3><div>Serum miR-943 levels were measured using RT-qPCR in 132 ACI patients and 135 healthy controls (HC). Diagnostic potential was evaluated by ROC analysis. Prognosis was assessed using 90-day mRS scores. Correlations with clinical parameters were analyzed using Pearson's/Spearman's tests. Multivariate logistic regression identified outcome predictors. Bioinformatic prediction and experimental validation (dual-luciferase, RIP assays) identified miR-943 targets. An OGD/R model in BV-2 microglia simulated ischemia-reperfusion. Effects of miR-943 inhibitors and si-BDNF on viability (CCK-8), apoptosis (flow cytometry), and inflammation (ELISA) were assessed.</div></div><div><h3>Results</h3><div>ACI patients exhibited significantly elevated serum miR-943 expression and reduced BDNF levels versus HC. miR-943 showed high diagnostic value (AUC = 0.909, sensitivity 85.6 %, specificity 83.7 %). MiR-943 expression correlated positively with SBP, DBP, FBG, Hcy, NIHSS score, and infarct volume. Patients with a poor prognosis exhibited significantly higher miR-943 expression levels. Multivariate analysis identified high expression of miR-943 as an independent risk factor for poor outcome. BDNF was validated as a direct miR-943 target gene. OGD/R increased miR-943 expression, suppressed proliferation, promoted apoptosis, and enhanced the expression of inflammatory factors in BV-2 cells. Inhibiting miR-943 expression reversed these detrimental effects, enhancing proliferation and reducing apoptosis/inflammation. Critically, BDNF knockdown abolished the protective effects of miR-943 inhibition.</div></div><div><h3>Conclusion</h3><div>Serum miR-943 is a promising diagnostic and prognostic biomarker for ACI. It exerts pathological effects by post-transcriptionally suppressing BDNF. Targeting the miR-943/BDNF axis represents a potential therapeutic strategy.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"775 ","pages":"Article 110667"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145407760","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}
Pub Date : 2026-01-01Epub Date: 2025-10-24DOI: 10.1016/j.abb.2025.110646
Jessica Georgina Filisola-Villaseñor , Samuel Álvarez-Almazán , Omar Granados-Portillo , Alicia Cruz-Jiménez , Gustavo G. Medina-Mendoza , Dan Israel Zavala-Vargas , J. Armando Luján-Montelongo , Edgar Morales-Ríos , Viridiana Olin-Sandoval
Ornithine decarboxylase (ODC) catalyzes the first step in the polyamines (PAs) biosynthesis pathway. These biomolecules are polycations that participate in many cellular processes. However, the non-physiological increase of human ODC (HsODC) expression can lead to the accumulation of PAs, and consequently, to the development of various types of cancer. Despite several inhibitors of HsODC have been proposed for cancer treatment, only the administration of difluoromethylornithine (DFMO) has been accepted as a drug for preventing the relapse of neuroblastoma. Thus, the search for other HsODC inhibitors continues. Previously, an alternative l-lysine decarboxylase activity was described in yeast and rat ODC. In this work, we expressed and purified the recombinant HsODC. Then, we kinetically characterized the ornithine and lysine decarboxylase (LDC) activities of the enzyme with Vmax/KM of 25.4 and 0.353 mL min−1 mg protein−1, respectively. Then, we selected 30 lysine-analogues from 300 candidate ligands for HsODC by molecular docking. The binding of these thirty molecules to HsODC was evaluated through thermal shift assays (TSA), obtaining nine binding molecules. Finally, the HsODC kinetic inhibition assays demonstrated that 7 compounds can inhibit this enzyme in vitro (with Ki ranging from 3.8 nM to 523.5 μM). With our results, we suggest these compounds as candidates to be explored as HsODC inhibitors in cell lines and in vivo models.
鸟氨酸脱羧酶（ODC）催化多胺（PAs）生物合成途径的第一步。这些生物分子是参与许多细胞过程的聚合体。然而，人类ODC （HsODC）表达的非生理性增加可导致PAs的积累，从而导致各种类型癌症的发生。尽管已经提出了几种HsODC抑制剂用于癌症治疗，但只有二氟甲基鸟氨酸（DFMO）被接受为预防神经母细胞瘤复发的药物。因此，对其他HsODC抑制剂的研究仍在继续。以前，在酵母和大鼠ODC中描述了另一种赖氨酸脱羧酶活性。在这项工作中，我们表达和纯化了重组HsODC。然后，我们对该酶的鸟氨酸和赖氨酸脱羧酶（LDC）活性进行了动力学表征，Vmax/KM分别为25.4和0.353 mL min - 1 mg protein - 1。然后，通过分子对接，从300个HsODC候选配体中筛选出30个赖氨酸类似物。通过热移法（TSA）评估了这30个分子与HsODC的结合，获得了9个结合分子。最后，HsODC动力学抑制实验表明，7种化合物对该酶具有体外抑制作用（Ki范围为3.8 nM ~ 523.5 μM）。根据我们的结果，我们建议这些化合物作为细胞系和体内模型中HsODC抑制剂的候选物进行探索。
{"title":"Characterization of the lysine decarboxylase activity of human ornithine decarboxylase and identification of lysine-like inhibitors","authors":"Jessica Georgina Filisola-Villaseñor , Samuel Álvarez-Almazán , Omar Granados-Portillo , Alicia Cruz-Jiménez , Gustavo G. Medina-Mendoza , Dan Israel Zavala-Vargas , J. Armando Luján-Montelongo , Edgar Morales-Ríos , Viridiana Olin-Sandoval","doi":"10.1016/j.abb.2025.110646","DOIUrl":"10.1016/j.abb.2025.110646","url":null,"abstract":"<div><div>Ornithine decarboxylase (ODC) catalyzes the first step in the polyamines (PAs) biosynthesis pathway. These biomolecules are polycations that participate in many cellular processes. However, the non-physiological increase of human ODC (HsODC) expression can lead to the accumulation of PAs, and consequently, to the development of various types of cancer. Despite several inhibitors of HsODC have been proposed for cancer treatment, only the administration of difluoromethylornithine (DFMO) has been accepted as a drug for preventing the relapse of neuroblastoma. Thus, the search for other HsODC inhibitors continues. Previously, an alternative <span>l</span>-lysine decarboxylase activity was described in yeast and rat ODC. In this work, we expressed and purified the recombinant HsODC. Then, we kinetically characterized the ornithine and lysine decarboxylase (LDC) activities of the enzyme with V<sub>max</sub>/K<sub>M</sub> of 25.4 and 0.353 mL min<sup>−1</sup> mg protein<sup>−1</sup>, respectively. Then, we selected 30 lysine-analogues from 300 candidate ligands for HsODC by molecular docking. The binding of these thirty molecules to HsODC was evaluated through thermal shift assays (TSA), obtaining nine binding molecules. Finally, the HsODC kinetic inhibition assays demonstrated that 7 compounds can inhibit this enzyme <em>in vitro</em> (with Ki ranging from 3.8 nM to 523.5 μM). With our results, we suggest these compounds as candidates to be explored as HsODC inhibitors in cell lines and <em>in vivo</em> models.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"775 ","pages":"Article 110646"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145414938","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}
Macrophages are homeostatic cells with remarkable phenotypic plasticity and key roles in inflammation. Macrophage lipoxygenases (LOX) oxygenate polyunsaturated fatty acids, producing oxylipins that are involved in inflammatory processes. Most insights into human LOX products and their roles in inflammation have been gained from animal models and human peripheral blood monocyte-derived macrophages (PB-MDMs). Human macrophage cell lines, although less explored, offer potential advantages due to their reduced culture complexity and variability. This study analyzes LOXes of human THP-1 monocyte-derived macrophages (THP-MDMs). LPS-polarized and IL-4-polarized macrophages (MLPS and MIL4) from THP-MDMs expressed the same LOX isoforms as polarized PB-MDMs. In both models, 5-LOX was similarly expressed in MLPS and MIL4, whereas 5-LOX activating protein was higher in MLPS. In contrast, 15-LOX1 and 15-LOX2 were predominantly expressed in MIL4. HPLC-MS/MS oxylipin analysis revealed that MIL4 from THP-MDMs and PB-MDMs produces similar metabolites of arachidonic, linoleic, eicosapentaenoic, and docosahexaenoic acids, including prostanoids, LOX, and cytochrome P450 monohydroxylated intermediates. Although precursors considered markers for resolvins and maresins pathways were identified, specialized pro-resolving mediators (SPMs) were not detected. These findings highlight the structural and functional similarities between PB-MDMs and THP-MDMs, positioning THP-MDMs as a promising model for studying macrophage LOX and oxylipin biosynthesis.
{"title":"Lipoxygenase expression and oxylipin analysis in human THP-1 monocyte-derived macrophages","authors":"Abramo Sofia , Mastrogiovanni Mauricio , Rubbo Homero , Gonzalez-Perilli Lucia","doi":"10.1016/j.abb.2025.110647","DOIUrl":"10.1016/j.abb.2025.110647","url":null,"abstract":"<div><div>Macrophages are homeostatic cells with remarkable phenotypic plasticity and key roles in inflammation. Macrophage lipoxygenases (LOX) oxygenate polyunsaturated fatty acids, producing oxylipins that are involved in inflammatory processes. Most insights into human LOX products and their roles in inflammation have been gained from animal models and human peripheral blood monocyte-derived macrophages (PB-MDMs). Human macrophage cell lines, although less explored, offer potential advantages due to their reduced culture complexity and variability. This study analyzes LOXes of human THP-1 monocyte-derived macrophages (THP-MDMs). LPS-polarized and IL-4-polarized macrophages (M<sub>LPS</sub> and M<sub>IL4</sub>) from THP-MDMs expressed the same LOX isoforms as polarized PB-MDMs. In both models, 5-LOX was similarly expressed in M<sub>LPS</sub> and M<sub>IL4</sub>, whereas 5-LOX activating protein was higher in M<sub>LPS</sub>. In contrast, 15-LOX1 and 15-LOX2 were predominantly expressed in M<sub>IL4</sub>. HPLC-MS/MS oxylipin analysis revealed that M<sub>IL4</sub> from THP-MDMs and PB-MDMs produces similar metabolites of arachidonic, linoleic, eicosapentaenoic, and docosahexaenoic acids, including prostanoids, LOX, and cytochrome P450 monohydroxylated intermediates. Although precursors considered markers for resolvins and maresins pathways were identified, specialized pro-resolving mediators (SPMs) were not detected. These findings highlight the structural and functional similarities between PB-MDMs and THP-MDMs, positioning THP-MDMs as a promising model for studying macrophage LOX and oxylipin biosynthesis.</div></div>","PeriodicalId":8174,"journal":{"name":"Archives of biochemistry and biophysics","volume":"775 ","pages":"Article 110647"},"PeriodicalIF":3.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145353433","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}
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