期刊：
Chemical Communications,2025年61(85):16498-16511 ISSN：1359-7345
通讯作者：
Xing, H
作者机构：
[Huang, Xiaowei; Lin, Yingwu] Univ South China, Sch Chem & Chem Engn, Hengyang 421001, Peoples R China.;[Xie, Wenxue; Huang, Xiaowei; Du, Jiajun; Li, Haokun; Xing, Hang] Hunan Univ, Inst Chem Biol & Nanomed, State Key Lab Chemo & Biosensing, Coll Chem & Chem Engn,Hunan Prov Key Lab Biomacrom, Changsha 410082, Peoples R China.;[Liu, Yuanyuan] Hunan Inst Drug Control, Changsha 410001, Hunan, Peoples R China.;[Xing, Hang] Hunan Univ Chongqing, Res Inst, Chongqing 401100, Peoples R China.
通讯机构：
[Xing, H ] H;Hunan Univ, Inst Chem Biol & Nanomed, State Key Lab Chemo & Biosensing, Coll Chem & Chem Engn,Hunan Prov Key Lab Biomacrom, Changsha 410082, Peoples R China.;Hunan Univ Chongqing, Res Inst, Chongqing 401100, Peoples R China.
摘要：
This highlight review article summarizes recent advances in employing HUH endonucleases as self-labeling protein tags for the sequence-specific covalent conjugation of unmodified ssDNA and examines their applications in cellular studies via engineered DNA-protein conjugates. We outline the structural basis and catalytic mechanism of the conserved HUH and Y motifs, which enable high selectivity, bioorthogonality, and robust conjugation under physiological conditions. Recent applications demonstrate the versatility of HUH-based DNA-protein conjugates in programmable cellular interface engineering, targeted therapeutic delivery, and enhancement of genome editing systems such as CRISPR-Cas. In the perspective section, we further highlight two emerging directions: computational tools such as the HUHgle platform for predictive substrate design, and directed evolution strategies extending HUH reactivity toward RNA substrates. Together, these advancements establish HUH endonucleases as powerful, programmable tools for generating DNA-protein conjugates that enable innovations in chemical biology, synthetic biology, and therapeutics.
期刊：
Molecular and Cellular Biochemistry,2025年480(4):2143-2157 ISSN：0300-8177
通讯作者：
Wang, J
作者机构：
[Jiang, Tingting] Univ South China, Hengyang Med Sch, Affiliated Nanhua Hosp, Dept Clin Lab, Hengyang 421000, Peoples R China.;[Zeng, Qun] Univ South China, Hengyang Med Sch, Dept Biochem & Mol Biol, Hengyang 421000, Peoples R China.;[Wang, Jing] Changsha Med Univ, Hunan Prov Key Lab Tradit Chinese Med Agr Biogenom, Changsha 410219, Peoples R China.;[Wang, Jing] Changsha Med Univ, Hunan Prov Univ Key Lab Fundamental & Clin Res Fun, Changsha 410219, Peoples R China.;[Wang, Jing] Changsha Med Univ, Clin Coll 1, Changsha 410219, Peoples R China.
通讯机构：
[Wang, J ] C;Changsha Med Univ, Hunan Prov Key Lab Tradit Chinese Med Agr Biogenom, Changsha 410219, Peoples R China.;Changsha Med Univ, Hunan Prov Univ Key Lab Fundamental & Clin Res Fun, Changsha 410219, Peoples R China.;Changsha Med Univ, Clin Coll 1, Changsha 410219, Peoples R China.
摘要：
FHL2 (Four-and-a-half LIM domain protein 2) is a crucial factor involved in cardiac morphogenesis, the process by which the heart develops its complex structure. It is expressed in various tissues during embryonic development, including the developing heart, and has been shown to play important roles in cell proliferation, differentiation, and migration. FHL2 interacts with multiple proteins to regulate cardiac development as a coactivator or a corepressor. It is involved in cardiac specification and determination of cell fate, cardiomyocyte growth, cardiac remodeling, myofibrillogenesis, and the regulation of HERG channels. Targeting FHL2 has therapeutic implications as it could improve cardiac function, control arrhythmias, alleviate heart failure, and maintain cardiac integrity in various pathological conditions. The identification of FHL2 as a signature gene in atrial fibrillation suggests its potential as a diagnostic marker and therapeutic target for this common arrhythmia.
摘要：
The major facilitator superfamily (MFS) type efflux pumps of Acinetobacter baumannii play important roles in antibiotic resistance. However, the molecular mechanism of these transporters remains poorly understood. To address the molecular basis of substrate polyspecificity mediated by multidrug MFS transporters, we compared the substrate binding modes of A. baumannii CraA with its well-studied homolog, Escherichia coli MdfA. MdfA and CraA share similar structural features, including a cavity accessible to drugs from the cytoplasm when these transporters adopt the inside-out conformation. This predominantly hydrophobic cavity contains several distinct titratable and hydrophilic residues. Through substitution analysis, we demonstrate that these polar residues within the CraA drug binding cavity contribute to the transport of all tested drugs, whereas mutations of hydrophobic residues result in altered drug recognition profiles. In addition to the known titratable residues E38 and D46, we identified E338 as the only titratable residue that plays a substrate-specific role, as it is required for efficient transport of norfloxacin, but not ethidium. Substitution of E338 with asparagine or glutamine changes substrate specificity, enabling specific recognition of phenicols and mitomycin C. Furthermore, we show that the aromaticity of Y42 is crucial for phenicol recognition, while general hydrophobicity at this position is critical for mitomycin C specificity. We propose that E338 and Y42 function as key substrate selectivity determinants in CraA. IMPORTANCE Multidrug efflux transporters of the major facilitator superfamily (MFS) are key contributors to antibiotic resistance, mediating the export of structurally diverse compounds across bacterial membranes. While homologous transporters such as Escherichia coli MdfA and Acinetobacter baumannii CraA share high structural similarity and overlapping substrate profiles, the molecular basis of their substrate specificity remains poorly understood. In this study, we show that structural homology among MFS transporters does not inherently imply mechanistic conservation, as species-specific variations can give rise to distinct substrate recognition profiles. Our findings reveal that CraA utilizes unique residues Y42 and E338 for substrate selectivity, while R124 and Y73 contribute to its transport activity. These findings enhance our understanding of efflux pump specificity and underscore the need to consider organism-specific features when targeting multidrug transporters in antimicrobial therapy.
The major facilitator superfamily (MFS) type efflux pumps of Acinetobacter baumannii play important roles in antibiotic resistance. However, the molecular mechanism of these transporters remains poorly understood. To address the molecular basis of substrate polyspecificity mediated by multidrug MFS transporters, we compared the substrate binding modes of A. baumannii CraA with its well-studied homolog, Escherichia coli MdfA. MdfA and CraA share similar structural features, including a cavity accessible to drugs from the cytoplasm when these transporters adopt the inside-out conformation. This predominantly hydrophobic cavity contains several distinct titratable and hydrophilic residues. Through substitution analysis, we demonstrate that these polar residues within the CraA drug binding cavity contribute to the transport of all tested drugs, whereas mutations of hydrophobic residues result in altered drug recognition profiles. In addition to the known titratable residues E38 and D46, we identified E338 as the only titratable residue that plays a substrate-specific role, as it is required for efficient transport of norfloxacin, but not ethidium. Substitution of E338 with asparagine or glutamine changes substrate specificity, enabling specific recognition of phenicols and mitomycin C. Furthermore, we show that the aromaticity of Y42 is crucial for phenicol recognition, while general hydrophobicity at this position is critical for mitomycin C specificity. We propose that E338 and Y42 function as key substrate selectivity determinants in CraA.
IMPORTANCE
Multidrug efflux transporters of the major facilitator superfamily (MFS) are key contributors to antibiotic resistance, mediating the export of structurally diverse compounds across bacterial membranes. While homologous transporters such as Escherichia coli MdfA and Acinetobacter baumannii CraA share high structural similarity and overlapping substrate profiles, the molecular basis of their substrate specificity remains poorly understood. In this study, we show that structural homology among MFS transporters does not inherently imply mechanistic conservation, as species-specific variations can give rise to distinct substrate recognition profiles. Our findings reveal that CraA utilizes unique residues Y42 and E338 for substrate selectivity, while R124 and Y73 contribute to its transport activity. These findings enhance our understanding of efflux pump specificity and underscore the need to consider organism-specific features when targeting multidrug transporters in antimicrobial therapy.
Multidrug efflux transporters of the major facilitator superfamily (MFS) are key contributors to antibiotic resistance, mediating the export of structurally diverse compounds across bacterial membranes. While homologous transporters such as Escherichia coli MdfA and Acinetobacter baumannii CraA share high structural similarity and overlapping substrate profiles, the molecular basis of their substrate specificity remains poorly understood. In this study, we show that structural homology among MFS transporters does not inherently imply mechanistic conservation, as species-specific variations can give rise to distinct substrate recognition profiles. Our findings reveal that CraA utilizes unique residues Y42 and E338 for substrate selectivity, while R124 and Y73 contribute to its transport activity. These findings enhance our understanding of efflux pump specificity and underscore the need to consider organism-specific features when targeting multidrug transporters in antimicrobial therapy.
Multidrug efflux transporters of the major facilitator superfamily (MFS) are key contributors to antibiotic resistance, mediating the export of structurally diverse compounds across bacterial membranes. While homologous transporters such as Escherichia coli MdfA and Acinetobacter baumannii CraA share high structural similarity and overlapping substrate profiles, the molecular basis of their substrate specificity remains poorly understood. In this study, we show that structural homology among MFS transporters does not inherently imply mechanistic conservation, as species-specific variations can give rise to distinct substrate recognition profiles. Our findings reveal that CraA utilizes unique residues Y42 and E338 for substrate selectivity, while R124 and Y73 contribute to its transport activity. These findings enhance our understanding of efflux pump specificity and underscore the need to consider organism-specific features when targeting multidrug transporters in antimicrobial therapy.
作者机构：
[Zhou, Daming; Qin, AJ; Zhang, Guiquan; Qin, Anjun] South China Univ Technol, Guangdong Prov Key Lab Luminescence Mol Aggregates, State Key Lab Luminescent Mat & Devices, Guangzhou 510640, Peoples R China.;[Tang, Ben Zhong] Chinese Univ Hong Kong, Shenzhen Inst Aggregate Sci & Technol, Sch Sci & Engn, Shenzhen CUHK Shenzhen, Shenzhen 518172, Peoples R China.;[Hu, Rong; Hu, R] Univ South China, Sch Chem & Chem Engn, Hengyang 421001, Peoples R China.
通讯机构：
[Qin, AJ ] S;[Tang, BZ ] C;[Hu, R ] U;South China Univ Technol, Guangdong Prov Key Lab Luminescence Mol Aggregates, State Key Lab Luminescent Mat & Devices, Guangzhou 510640, Peoples R China.;Chinese Univ Hong Kong, Shenzhen Inst Aggregate Sci & Technol, Sch Sci & Engn, Shenzhen CUHK Shenzhen, Shenzhen 518172, Peoples R China.
摘要：
Fluorescence imaging technology is playing increasing roles in modern personalized and precision medicine. Thanks to their excellent photophysical properties, organic luminogens featuring aggregation-induced emission (AIE) characteristics (AIEgens) have attracted considerable attention over the past two decades. Because of their superior biocompatibility, ease of processing and functionalization, excellent water solubility, high responsiveness, and exceptional signal-to-noise ratio (SNR) for biotargets, AIE bioconjugates, formed by covalently linking AIEgens with biomolecules, have emerged as an ideal candidate for bioapplications. In this review, we summarize the progress in preparation, properties, and application of AIE bioconjugates in the last five years. Moreover, the challenges and opportunities of AIE bioconjugates are also briefly discussed.
摘要：
The rapid and selective identification of microorganisms is of great significance for clinical therapy applications. To develop high performance probes for microbe determination, we systemically constructed series aggregation-induced emission (AIE) luminogens by modulating the structural planarity, the basicity of functional group, the length of linker moiety and the hydrophobicity based on our previous work. The detail structure-property relationship study based on experimental and theoretical observation revealed that: i) the planar skeleton is essential for probe insertion towards the cell wall via van n der Waals' force. ii) the basic function group enable the anchoring on the membrane by binding with acidic biomolecules. iii) the shortened alkyl chain is in favor of the efficient binding of basic groups with microbes and endows the desirable hydrophobicity. Based on the developed probes, the successful detection of the pathogens in clinic samples was achieved in highly sensitive and simple way. This work provides a reliable strategy for designing intelligent luminogens for microorganism discrimination and identification in efficient and sensitive way for in vitro diagnosis applications, especially point-of-care testing (POCT).
The rapid and selective identification of microorganisms is of great significance for clinical therapy applications. To develop high performance probes for microbe determination, we systemically constructed series aggregation-induced emission (AIE) luminogens by modulating the structural planarity, the basicity of functional group, the length of linker moiety and the hydrophobicity based on our previous work. The detail structure-property relationship study based on experimental and theoretical observation revealed that: i) the planar skeleton is essential for probe insertion towards the cell wall via van n der Waals' force. ii) the basic function group enable the anchoring on the membrane by binding with acidic biomolecules. iii) the shortened alkyl chain is in favor of the efficient binding of basic groups with microbes and endows the desirable hydrophobicity. Based on the developed probes, the successful detection of the pathogens in clinic samples was achieved in highly sensitive and simple way. This work provides a reliable strategy for designing intelligent luminogens for microorganism discrimination and identification in efficient and sensitive way for in vitro diagnosis applications, especially point-of-care testing (POCT).
期刊：
PHYSICAL CHEMISTRY CHEMICAL PHYSICS,2025年27(5):2828-2833 ISSN：1463-9076
通讯作者：
Liu, Xichun;Lin, YW
作者机构：
[Han, Hui; Pan, Aiqun; Lin, Ying-Wu; Liu, Xichun; Liu, XC] Univ South China, Sch Chem & Chem Engn, Hengyang 421001, Peoples R China.;[Gao, Shu-Qin; Lin, Ying-Wu] Univ South China, Lab Prot Struct & Funct, Hengyang 421001, Peoples R China.
通讯机构：
[Lin, YW ; Liu, XC] U;Univ South China, Sch Chem & Chem Engn, Hengyang 421001, Peoples R China.;Univ South China, Lab Prot Struct & Funct, Hengyang 421001, Peoples R China.
摘要：
Globin X is a newly discovered member of the globin family, while its structure and function are not fully understood. In this study, we performed protein modelling studies using Alphafold3 and molecular dynamics simulations, which suggested that the protein adopts a typical globin fold, with the formation of a potential disulfide bond of Cys65 and Cys141. To elucidate the role of this unique disulfide in protein structure and stability, we constructed a double mutant of C65S/C141S by mutating the two cysteine residues to serine. As suggested by protein mass, ultraviolet-visible (UV-Vis) and circular dichroism (CD) spectroscopy analyses, the potential disulfide bond has minimal effect on the overall protein structure, but its absence reduces the protein stability. Electron paramagnetic resonance (EPR) analysis also revealed an increase in the proportion of high-spin state heme iron, which accelerates the rate of heme degradation in reaction with H(2)O(2). This study highlights the critical role of the Cys65-Cys141 in maintaining the stability of globin X and the bis-His heme coordination state, providing insights into the structure-function relationship of the newly discovered globin.
摘要：
The sensitive detection of the radioactive thorium (Th) ion with an oxidation state of +4 (Th(4+)) is of great significance for environmental protection and life safety. In this study, five fluorescence sensors with regulated donor-acceptor (D-A) interactions were constructed for Th(4+) detection based on intramolecular charge transfer and aggregation-induced emission mechanisms. Among the developed sensors, TPE-D bearing electron-deficient π-bridge and weak D-A interactions presented ratiometric fluorescence detection behavior toward Th(4+) in aqueous solution due to its aggregation-induced emission characteristics and unique D-A-D structures. Moreover, TPE-D showed excellent selectivity and sensitivity for Th(4+) detection, and the detection limit was as low as 8.1 × 10(-8) M. The sensing mechanism observation revealed that Th(4+) could coordinate with the hydroxyl, imine, and carbonyl groups of TPE-D accompanied by an electron transfer process. In addition, TPE-D could selectively be enriched in the lysosome. Both the detection of Th(4+) in the lysosome and liver of mice and zebrafish were realized based on this strategy, and a mobile-assisted detection approach toward Th(4+) in actual water samples was also established with high sensitivity. This is the first report for Th(4+) detection in organelles and organs, which provides a great significance and reliable strategy for radionuclide toxicology detection and analysis applications.
摘要：
Copper serves as a crucial trace element in various biological systems. Copper ions form complexes with different ligands, amplifying reactive oxygen species (ROS) levels and promoting intracellular ROS accumulation in multiple cancer cell types. In this study, a copper(II) complex, dichlorido[4-(5-bromothiazol-2-yl)-2,2'-bipyridine] copper(II) (Cu1), was synthesized using a terpyridine derivative as the ligand. Its structural configuration was confirmed through mass spectrometry and single-crystal X-ray diffraction analysis. The in vitro anticancer efficacy and mechanisms of Cu1 against B16-F10 cells were systematically examined using an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, apoptosis analysis, comet assay, invasion assay, wound healing assay, western blotting, and 3D tumor spheroid models. The findings demonstrated that Cu1 exhibited significant cytotoxicity toward B16-F10 cells and induced dose-dependent apoptosis. Additionally, Cu1 stimulated ROS generation and accumulation in B16-F10 melanoma cells, activating the NLRP3 (NOD-like receptor family pyrin domain-containing 3) inflammasome and caspase-1. Activated caspase-1 specifically cleaved gasdermin D (GSDMD), releasing its N-terminal domain (GSDMD-N), which formed pores in the cell membrane, leading to pyroptosis and DNA double-strand breaks. Cu1 also exhibited significant growth-inhibitory effects in 3D tumor spheroid models, underscoring its potential applicability in simulating in vivo tumor microenvironments.
期刊：
Journal of Colloid and Interface Science,2025年692:137529 ISSN：0021-9797
通讯作者：
Xue, FQ;Liu, Xiaolong;Wang, PY
作者机构：
[Zhang, Ziqian; Xue, Fangqin; Xu, Chao; He, Liangzhen; Xue, FQ] Fuzhou Univ, Affiliated Prov Hosp, Fujian Prov Hosp, Fujian Med Univ,Dept Gastrointestinal Surg,Shengli, Fuzhou 350001, Peoples R China.;[Ying, Yunfei; Wang, Peiyuan; Zhang, Ziqian; Liu, Xiaolong; Dang, Yongying; Li, Siyaqi; Liu, XL] Chinese Acad Sci, Fujian Inst Res Struct Matter, Fuzhou 350002, Peoples R China.;[Liu, Xiaolong; Wang, Peiyuan] Fujian Med Univ, United Innovat Mengchao Hepatobiliary Technol Key, Mengchao Hepatobiliary Hosp, Fuzhou 350025, Peoples R China.;[Ying, Yunfei] Univ South China, Sch Basic Med, Dept Biochem & Mol Biol, Hengyang 421001, Peoples R China.;[Li, Siyaqi] Fujian Agr & Forestry Univ, Coll Life Sci, Fuzhou 350002, Peoples R China.
通讯机构：
[Wang, PY ; Liu, XL] C;[Xue, FQ ] F;Fuzhou Univ, Affiliated Prov Hosp, Fujian Prov Hosp, Fujian Med Univ,Dept Gastrointestinal Surg,Shengli, Fuzhou 350001, Peoples R China.;Chinese Acad Sci, Fujian Inst Res Struct Matter, Fuzhou 350002, Peoples R China.
关键词：
Fluorescence imaging;Multi ion synergistic;PTT enhanced CDT;Tumor microenvironment;Tumor surgery
摘要：
Colon cancer, characterized by its high incidence and mortality rates, continues to present a significant challenge in cancer treatment. To address this, we present a novel ZnCe based nanocarrier featuring stacked mesopores and rough surface, indocyanine Green (ICG) is encapsulated within these mesopores (ZnCe&ICG). This innovative nanoplatform demonstrates effective accumulation in tumor regions and can be triggered to generate efficacious reactive oxygen species (ROS) in the weakly acidic and high H 2 O 2 conditions typical of tumor microenvironments. Enhanced fluorescent imaging using improved tumor-to-background ratio has proven effective in precisely delineating tumor margins from surrounding healthy tissue. With the guidance of this second near-infrared region (NIR II, 1000–1700 nm) fluorescence imaging technique, tumors are completely excised, resulting in negligible instances of in situ recurrence or metastasis observed 30 days following surgery. Notably, under 808 nm laser irradiation, the nanoplatform exhibits a high photothermal conversion efficiency, leading to localized heating that further amplifies ROS production via multi ion synergetic catalysis for tumor cell killing. These results underscore the potential of tumor microenvironment-responsive ZnCe-based nanocomposite as a fluorescence imaging contrast agent and chemodynamic agent for cancer treatment, particularly when combined with NIR light activation.
Colon cancer, characterized by its high incidence and mortality rates, continues to present a significant challenge in cancer treatment. To address this, we present a novel ZnCe based nanocarrier featuring stacked mesopores and rough surface, indocyanine Green (ICG) is encapsulated within these mesopores (ZnCe&ICG). This innovative nanoplatform demonstrates effective accumulation in tumor regions and can be triggered to generate efficacious reactive oxygen species (ROS) in the weakly acidic and high H 2 O 2 conditions typical of tumor microenvironments. Enhanced fluorescent imaging using improved tumor-to-background ratio has proven effective in precisely delineating tumor margins from surrounding healthy tissue. With the guidance of this second near-infrared region (NIR II, 1000–1700 nm) fluorescence imaging technique, tumors are completely excised, resulting in negligible instances of in situ recurrence or metastasis observed 30 days following surgery. Notably, under 808 nm laser irradiation, the nanoplatform exhibits a high photothermal conversion efficiency, leading to localized heating that further amplifies ROS production via multi ion synergetic catalysis for tumor cell killing. These results underscore the potential of tumor microenvironment-responsive ZnCe-based nanocomposite as a fluorescence imaging contrast agent and chemodynamic agent for cancer treatment, particularly when combined with NIR light activation.
摘要：
Given the severe environmental and health risks posed by heavy metal pollutants such as mercury (Hg), cadmium (Cd), lead (Pb), chromium (Cr), and arsenic (As), the development of highly efficient and selective capture materials is critically important.This study explores the molecular structural characteristics of complexes formed by substituting the central palladium atom in nitrogen-heterocyclic carbene complexes with mercury (Hg), cadmium (Cd), lead (Pb), chromium (Cr), and arsenic (As). For all the complexes, the simulated infrared and ultraviolet-visible spectra, Wiberg bond indices, as well as the binding ability of nitrogen-heterocyclic carbene ligands to the central atom were assessed. The results show that the complexes obtained when the central atom is replaced by Hg, Cd, Pb, Cr, or As are stable and display unique spectral features. Significantly, nitrogen-heterocyclic carbene has the strongest binding affinity for Pd. The complexes of chromium substitutes possess relatively high chemical stability. These findings will offer important references for future pollution control and support the specific capture of the five studied heavy metals, and enrichment of Cr using nitrogen-heterocyclic carbene ligands.
摘要：
Aptamers have recently become novel probes for biosensors because of their good biocompatibility, strong specificity, and high sensitivity. Biosensors based on peptides or nucleic acid aptamers are used in implantable and wearable devices owing to their ease of synthesis and economic efficiency. Simultaneously, amphoteric ionic peptides are being explored as antifouling layers for biosensors resistant to interference from extraneous proteins in serum. Thus, this paper reviews recently developed aptamer-based biosensors and introduces peptide- and nucleic acid-based biosensors, while focusing on the three primary classes of biosensors: electrochemical sensors, fluorescent or colorimetric biosensors, and electroluminescent sensors. Furthermore, we summarize their general construction strategies, describe specific electrochemical sensors that use peptides as an antipollution layer, and elucidate their advantages.
摘要：
Chloroanilines represent a class of persistent and highly toxic environmental pollutants, posing significant challenges for green remediation strategies. While P450BM3 monooxygenases are renowned for their ability to catalyze the monooxidation of inert C-H bonds, costly NAD(P)H and complex electron transport systems required for P450BM3 catalysis limit their practical applications. This study pioneers the development of innovative artificial biocatalysts by strategically engineering the active site of P450BM3. Specifically, the substitution of the highly conserved threonine 268 with aspartic acid effectively induces peroxygenase activity, allowing for enhanced catalytic efficiency. Remarkably, the engineered P450BM3 mutants achieved degradation rates of 98.38–99.18 % for five chloroanilines (4-chloroaniline, 2-chloroaniline, 2,4-dichloroaniline, 3,4-dichloroaniline, and 3,5-dichloroaniline) in just 10–15 min, all without the need for NAD(P)H or dual-functional small molecules. Comprehensive degradation mechanism analysis via UPLC-MS corroborated the remarkable performance of these biocatalysts. This research not only demonstrates a novel approach for engineering P450 monooxygenases to exhibit peroxygenase activity but also significantly broadens their potential applications in synthetic chemistry and synthetic biology, paving the way for greener and more sustainable remediation technologies.
Chloroanilines represent a class of persistent and highly toxic environmental pollutants, posing significant challenges for green remediation strategies. While P450BM3 monooxygenases are renowned for their ability to catalyze the monooxidation of inert C-H bonds, costly NAD(P)H and complex electron transport systems required for P450BM3 catalysis limit their practical applications. This study pioneers the development of innovative artificial biocatalysts by strategically engineering the active site of P450BM3. Specifically, the substitution of the highly conserved threonine 268 with aspartic acid effectively induces peroxygenase activity, allowing for enhanced catalytic efficiency. Remarkably, the engineered P450BM3 mutants achieved degradation rates of 98.38–99.18 % for five chloroanilines (4-chloroaniline, 2-chloroaniline, 2,4-dichloroaniline, 3,4-dichloroaniline, and 3,5-dichloroaniline) in just 10–15 min, all without the need for NAD(P)H or dual-functional small molecules. Comprehensive degradation mechanism analysis via UPLC-MS corroborated the remarkable performance of these biocatalysts. This research not only demonstrates a novel approach for engineering P450 monooxygenases to exhibit peroxygenase activity but also significantly broadens their potential applications in synthetic chemistry and synthetic biology, paving the way for greener and more sustainable remediation technologies.
摘要：
An activatable photosensitizer with a reaction‐tunable donor/acceptor push‐pull electronic effect is designed and developed, which shows high sensitivity toward nitric oxide (NO) NO with the formation of triazole segment. Moreover, the turn‐on PDT behavior and endoplasmic reticulum (ER) targeting ability enable the developed photosensitizer good inhibition effect toward tumor cells both in vitro and in vivo, which presents high promise for precise tumor therapy in practical. Abstract Photodynamic therapy (PDT) is a powerful strategy for tumor therapy with noninvasiveness and desirable efficacy. However, the phototoxicity of photosensitizer after the post‐PDT is the major obstacle limiting the clinic applications. Herein, a nitric oxide (NO)‐activatable photosensitizer is reported with turn‐on PDT behavior and endoplasmic reticulum (ER) targeting ability for precise tumor therapy. Four o‐thiophenediamine derivatives with reaction‐tunable donor/acceptor push‐pull electronic effect are established, and the systematic structure and property relationship observation reveals the following features: 1) the reactivity against NO can be improved by enhancing the electron density and further facilitated upon photo‐irradiation. 2) the reactivity with NO enables the improved intramolecular charge transfer process with the evoking of photosensitizing effect. 3) only o‐thiophenediamine derivative with ER enrichment behavior exhibited cancer cell ablation effect compared to photosensitizers localized in lysosome and lipid droplet. Thus, the efficient inhibition of cancer cells both in vitro and in vivo is realized based on the photo‐controlled PDT strategy. This work provides more insights into developing promising activatable photosensitizers for advanced therapy based on tumor microenvironment trigger.
作者机构：
[Zhang, Yuexin; Li, Ye; Yin, Mingxue; Li, Chunquan; Liu, Liyuan; Zhang, Guorui; Zhang, Jianing; Song, Chao] Univ South China, Affiliated Hosp 1, Hengyang 421001, Hunan, Peoples R China.;[Zhang, Yuexin; Li, Ye; Yin, Mingxue; Li, Chunquan; Liu, Liyuan; Zhang, Guorui; Zhang, Jianing; Song, Chao] Univ South China, Hengyang Med Sch, Natl Hlth Commiss Key Lab Birth Defect Res & Preve, Hengyang 421001, Hunan, Peoples R China.;[Zhang, Yuexin; Li, Ye; Yin, Mingxue; Li, Chunquan; Liu, Liyuan; Zhang, Guorui; Zhang, Jianing; Song, Chao] Univ South China, Hunan Prov Key Lab Multiomics&Artificial Intellige, Hengyang 421001, Hunan, Peoples R China.;[Li, Ye; Yin, Mingxue; Li, Chunquan; Liu, Liyuan; Zhang, Guorui] Univ South China, Sch Basic Med Sci, Hengyang Med Sch, Dept Biochem & Mol Biol, Hengyang 421001, Hunan, Peoples R China.;[Li, Chunquan; Song, Chao] Univ South China, Sch Comp, Hengyang 421001, Hunan, Peoples R China.
通讯机构：
[Li, CQ ] U;[Guo, MZ ] B;Univ South China, Affiliated Hosp 1, Hengyang 421001, Hunan, Peoples R China.;Univ South China, Hengyang Med Sch, Natl Hlth Commiss Key Lab Birth Defect Res & Preve, Hengyang 421001, Hunan, Peoples R China.;Univ South China, Hunan Prov Key Lab Multiomics&Artificial Intellige, Hengyang 421001, Hunan, Peoples R China.
摘要：
It is challenging to identify regulatory transcriptional regulators (TRs), which control gene expression via regulatory elements and epigenomic signals, in context-specific studies on the onset and progression of diseases. The use of large-scale multi-omics epigenomic data enables the representation of the complex epigenomic patterns of control of the regulatory elements and the regulators. Herein, we propose Transcription Regulator Activity Prediction Tool (TRAPT), a multi-modality deep learning framework, which infers regulator activity by learning and integrating the regulatory potentials of target gene cis-regulatory elements and genome-wide binding sites. The results of experiments on 570 TR-related datasets show that TRAPT outperformed state-of-the-art methods in predicting the TRs, especially in terms of forecasting transcription co-factors and chromatin regulators. Moreover, we successfully identify key TRs associated with diseases, genetic variations, cell-fate decisions, and tissues. Our method provides an innovative perspective on identifying TRs by using epigenomic data. Here, the authors propose Transcription Regulator Activity Prediction Tool (TRAPT), a multi-modality deep learning framework, which infers regulator activity by learning and integrating the regulatory potentials of target gene cis-regulatory elements and genome-wide binding sites.
期刊：
International Journal of Biological Macromolecules,2025年309(Pt 1):142768 ISSN：0141-8130
通讯作者：
Xu, JK
作者机构：
[Zhao, Sijia; Li, Dong; Xu, Jiakun; Wang, Fang; Wang, Jinghan] Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, State Key Lab Mariculture Biobreeding & Sustainabl, Key Lab Sustainable Dev Polar Fisheries,Minist Agr, Qingdao 266071, Peoples R China.;[Zhao, Sijia; Wang, Jinghan] Ocean Univ China, Coll Food Sci & Engn, Qingdao Marine Sci & Technol Ctr, Lab Marine Drugs & Bioprod, Qingdao 266003, Peoples R China.;[Shoji, Osami] Nagoya Univ, Grad Sch Sci, Dept Chem, Furo Cho,Chikusa Ku, Nagoya 4648602, Japan.;[Lin, Yingwu] Univ South China, Sch Chem & Chem Engn, Hengyang 421001, Peoples R China.
通讯机构：
[Xu, JK ] C;Chinese Acad Fishery Sci, Yellow Sea Fisheries Res Inst, State Key Lab Mariculture Biobreeding & Sustainabl, Key Lab Sustainable Dev Polar Fisheries,Minist Agr, Qingdao 266071, Peoples R China.
关键词：
Lignin;O-demethylation;P450BM3
摘要：
The enzymatic demethylation of aromatic compounds presents a major challenge in the valorization of lignin. The main goal was to develop an efficient artificial peroxygenase system combining engineered P450BM3 with AldO (sugar alcohol oxidase) and DFSM (dual function small molecule) for the regioselective O -demethylation of lignin-derived aromatic ethers. P450BM3 serves as a versatile biocatalyst, and its engineered variants demonstrate expanded substrate promiscuity toward non-native substrates. AldO, served as the H 2 O 2 in situ generation system. The DFSM, a rationally designed catalytic auxiliary, facilitates precise control of enzymatic reactions and enhances the efficiency of O -demethylation. We hypothesize that by combining P450BM3 with AldO and DFSM, we can better control the generation of H 2 O 2 and direct the enzymatic system toward efficient O -demethylation. The engineered P450BM3 F87A/V78A/T268D/A328F mutant achieved a TON of 1895 ± 4 for guaiacol, more than double that of the native P450BM3/H 2 O 2 system (TON = 872 ± 7). Moreover, the F87A/T268D mutant efficiently catalyzed double-demethylation of syringol, achieving the highest turnover number (TON) of 483 ± 7. This DFSM-assisted P450BM3/AldO system represents a significant advancement in the biocatalytic degradation of lignin and offers a cost-effective and scalable alternative to traditional NADPH-dependent P450 monooxygenases. Our findings open new pathways for sustainable biotechnological applications in lignin valorization and aromatic compound catabolism.
The enzymatic demethylation of aromatic compounds presents a major challenge in the valorization of lignin. The main goal was to develop an efficient artificial peroxygenase system combining engineered P450BM3 with AldO (sugar alcohol oxidase) and DFSM (dual function small molecule) for the regioselective O -demethylation of lignin-derived aromatic ethers. P450BM3 serves as a versatile biocatalyst, and its engineered variants demonstrate expanded substrate promiscuity toward non-native substrates. AldO, served as the H 2 O 2 in situ generation system. The DFSM, a rationally designed catalytic auxiliary, facilitates precise control of enzymatic reactions and enhances the efficiency of O -demethylation. We hypothesize that by combining P450BM3 with AldO and DFSM, we can better control the generation of H 2 O 2 and direct the enzymatic system toward efficient O -demethylation. The engineered P450BM3 F87A/V78A/T268D/A328F mutant achieved a TON of 1895 ± 4 for guaiacol, more than double that of the native P450BM3/H 2 O 2 system (TON = 872 ± 7). Moreover, the F87A/T268D mutant efficiently catalyzed double-demethylation of syringol, achieving the highest turnover number (TON) of 483 ± 7. This DFSM-assisted P450BM3/AldO system represents a significant advancement in the biocatalytic degradation of lignin and offers a cost-effective and scalable alternative to traditional NADPH-dependent P450 monooxygenases. Our findings open new pathways for sustainable biotechnological applications in lignin valorization and aromatic compound catabolism.
通讯机构：
[Liao, S ] U;Univ South China, Sch Chem & Chem Engn, Hengyang 421001, Hunan, Peoples R China.
摘要：
By utilizing carboxylated multiwalled carbon nanotubes (MWCNT-COOH) to strengthen the interaction between the electrode and the analytes and improve the conductivity of the composite material and in conjunction with the superior catalytic properties of copper-based metal-organic framework (MOFs), a novel electrochemical sensor was fabricated from a Cu-MOF/MWCNT-COOH composite, specifically designed for the simultaneous and distinct detection of ascorbic acid (AA) and dopamine (DA). Electrochemical analyses were conducted on the innovative Cu-MOF/MWCNT-COOH electrode through both CV and DPV, revealing unique electrochemical behaviors for AA and DA. The sensor not only showed exceptional electrocatalytic properties but also distinguished itself by its broad dynamic response ranges, covering concentrations from 3 to 1800 μM for AA and from 2 to 180 μM for DA, with detection limits (S/N = 3) of 3.00 μM for AA and 0.32 μM for DA. Furthermore, this electrochemical detection platform exhibited robust reproducibility and selectivity. Examinations of serum samples yielded the recovery rates of AA and DA which were 101.9% and 102.1%, respectively, confirming the sensor's capability to perform reliably under varied biological conditions. The findings confirm the sensor's potential of the proposed method for the simultaneous, sensitive, and reliable detection of AA and DA. In conclusion, the electrochemical sensor has a promising potential for practical applications.
摘要：
Cardiac fibrosis is a maladaptive response in which excessive extracellular matrix (ECM) deposition stiffens the myocardium and compromises systolic and diastolic function. Reactive oxygen species (ROS) sit at the hub of this process, acting as initiators and amplifiers of four key pro-fibrotic signaling cascades, including transforming growth factor-beta (TGF-β), mitogen-activated protein kinase (MAPK), nuclear factor kappa B (NF-κB), and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt). Crosstalk among these pathways forms a self-sustaining network that perpetuates fibroblast activation, ECM synthesis and inflammatory cytokine release. Targeting ROS generation, scavenging downstream oxidants, or selectively interrupting these signaling nodes therefore represents a rational strategy for attenuating myocardial fibrosis and restoring cardiac performance.
Cardiac fibrosis is a maladaptive response in which excessive extracellular matrix (ECM) deposition stiffens the myocardium and compromises systolic and diastolic function. Reactive oxygen species (ROS) sit at the hub of this process, acting as initiators and amplifiers of four key pro-fibrotic signaling cascades, including transforming growth factor-beta (TGF-β), mitogen-activated protein kinase (MAPK), nuclear factor kappa B (NF-κB), and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt). Crosstalk among these pathways forms a self-sustaining network that perpetuates fibroblast activation, ECM synthesis and inflammatory cytokine release. Targeting ROS generation, scavenging downstream oxidants, or selectively interrupting these signaling nodes therefore represents a rational strategy for attenuating myocardial fibrosis and restoring cardiac performance.
摘要：
The development of enzyme activity analysis methods is critical for precise and rapid assessments of enzyme activity levels within biological systems, facilitating a more profound comprehension of physiological functions and disease mechanisms. Alkaline phosphatase (ALP) participates in various physiological processes involving phosphate ester hydrolysis. Altered ALP activity levels are often indicative of different diseases, underscoring the necessity for accurate ALP activity determination in medical diagnostics. This study innovatively applies turbidity as a physical variable, proposing a turbidimetric sensor based on an enhanced ammonium molybdate reagent for phosphate analysis. By integrating this with the ALP substrate p-nitrophenyl phosphate, a turbidimetric sensor was devised and employed for ALP activity analysis. The proposed turbidimetric sensor demonstrated high sensitivity both for phosphate (0.18 μmol/L) and ALP activity (0.03 mU/mL) assay. In practical applications, this turbidimetric sensor has been effectively employed to detect ALP activity in mouse feces, showcasing its potential for auxiliary diagnosis of inflammatory bowel disease. Significantly, this novel turbidity-based approach offers not only swift and straightforward procedures but also remarkable portability and cost-efficiency. Requiring solely a handheld turbidimeter and eliminating the need for bulky instruments, this approach holds significant potential for point-of-care testing applications.
The development of enzyme activity analysis methods is critical for precise and rapid assessments of enzyme activity levels within biological systems, facilitating a more profound comprehension of physiological functions and disease mechanisms. Alkaline phosphatase (ALP) participates in various physiological processes involving phosphate ester hydrolysis. Altered ALP activity levels are often indicative of different diseases, underscoring the necessity for accurate ALP activity determination in medical diagnostics. This study innovatively applies turbidity as a physical variable, proposing a turbidimetric sensor based on an enhanced ammonium molybdate reagent for phosphate analysis. By integrating this with the ALP substrate p-nitrophenyl phosphate, a turbidimetric sensor was devised and employed for ALP activity analysis. The proposed turbidimetric sensor demonstrated high sensitivity both for phosphate (0.18 μmol/L) and ALP activity (0.03 mU/mL) assay. In practical applications, this turbidimetric sensor has been effectively employed to detect ALP activity in mouse feces, showcasing its potential for auxiliary diagnosis of inflammatory bowel disease. Significantly, this novel turbidity-based approach offers not only swift and straightforward procedures but also remarkable portability and cost-efficiency. Requiring solely a handheld turbidimeter and eliminating the need for bulky instruments, this approach holds significant potential for point-of-care testing applications.
摘要：
PB1-like phages belong to Pbunavirus and are widespread in various environments. This group of phages is a promising candidate for treating human or animal infectious diseases caused by antibiotic-resistant P. aeruginosa. The lipopolysaccharide (LPS) has been identified as the receptor of different PB1-like phages, while little is known about the receptor-binding proteins (RBPs) of these phages. We constructed the tail fiber protein (gp50) of a PB1-like phage, PHW2, and its C- or N-terminus truncation variants to identify its role during the phage infection. The anti-gp50((453-964)) antibody showed a similar effect to the antibody against gp50 in blocking the phage infection. The protein competition and cell binding assays showed that the gp50((1-451)) doesn't exhibit an effect on the adsorption of the host cells. These results indicated that the C-terminus of gp50 is the essential region that mediates phage PHW2 adsorption and infection.
摘要：
Spatially resolved transcriptomics technologies potentially provide the extra spatial position information and tissue image to better inferspatial cell-cell interactions (CCIs) in processes such as tissue homeostasis, development, and disease progression. However, methods for effectively integrating spatial multimodal data to infer CCIs are still lacking. Here, the authors propose a deep learning method for integrating features through co-convolution, called SpaGraphCCI, to effectively integrate data from different modalities of SRT by projecting gene expression and image feature into a low-dimensional space. SpaGraphCCI can achieve significant performance on datasets from multiple platforms including single-cell resolution datasets (AUC reaches 0.860-0.907) and spot resolution datasets (AUC ranges from 0.880 to 0.965). SpaGraphCCI shows better performance by comparing with the existing deep learning-based spatial cell communication inference methods. SpaGraphCCI is robust to high noise and can effectively improve the inference of CCIs. We test on a human breast cancer dataset and show that SpaGraphCCI can not only identify proximal cell communication but also infer new distal interactions. In summary, SpaGraphCCI provides a practical tool that enables researchers to decipher spatially resolved cell-cell communication based on spatial transcriptome data.
