aBIOTECH
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2025
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IF 5
Zhang Wenyang,Wang Yanjiao,Li Wenyan,Wu Shaowen,Chen Yuanyuan,Ye Mingyang,Huang Wenjie,Fernie Alisdair R.,Yan Shijuan
Protein post-translational modifications such as phosphorylation and S -nitrosylation regulate protein functions and cellular programs in eukaryotes. Moreover, extensive evidence suggests crosstalk between these modifications. However, we lack a comprehensive method for the simultaneous detection and analysis of multiple post-translational modifications. Here, we present an optimized workflow that identifies phosphorylation and S -nitrosylation sites using a novel phosphate affinity tag switch technique. Validation with model proteins and complex biological samples confirmed the high sensitivity, coverage, and reproducibility of this method. Applying this method to Arabidopsis thaliana seedlings revealed 12,552 phosphorylation sites and 6,108 S -nitrosylation sites, representing the largest single-study dataset of S -nitrosylation sites to date. This approach enhances our understanding of post-translational modification dynamics in plant signaling, stress responses, and metabolism.
研究领域:
植物生物学
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生物化学
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分子生物学
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蛋白质组学
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DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2023
·
IF 4.1
Hongmei Yang,Linxu Wang,Zhaoyang Xie,Simeng Shao,Yi Wu,Weiyin Xu,Bin Gu,Bo Wang
Comprehensive sulfur-nitrosylation (SNO) proteome coverage in complex biological systems remains challenging as a result of the low level of endogenous S-nitrosylation and its chemical instability. Herein, we optimized the synthesis route of SNOTRAP (SNO trapping by triaryl phosphine) probe and the proteomics pipeline (including preventing over-alkylation, sample washing, trypsin digestion). Preventing overalkylation was found to be the key factor resulting in a higher number of identified SNO proteins by evaluating various experimental conditions. With the improved SNOTRAP-based proteomic pipeline, we achieved an improvement of ∼10-fold on identification efficiency, and identified 1181 SNO proteins (1714 SNO sites) in mouse brain, representing the largest repository of endogenous S-nitrosylation. Moreover, we identified the consensus motif of SNO sites, suggesting the correlation with local hydrophobicity, acid-base catalysis, and the surrounding secondary structures for modification of specific cysteines by NO. Collectively, we provide a universal pipeline for the high-coverage identification of low-abundance SNO proteins with high enrichment efficiency, high specificity (98%), good reproducibility, and easy implementation, contributing to the elucidation of the mechanism(s) of nitrosative stress in multiple diseases.
研究领域:
生物化学
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蛋白质组学
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医学
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DOI
PubMed
高影响因子
CANCER SCIENCE
·
2020
·Vol. 111
·期 10
·3802-3812
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IF 4.966
Xiaolong Zhang,Qinli Ruan,Yiqian Zhai,Dandan Lu,Chen Li,Yahan Fu,Zihui Zheng,Ying Song,Jun Guo
Baicalein, a flavonoid phytochemical, has been shown to be effective as an anti-metastatic agent for various cancers, especially for non-small-cell lung cancer (NSCLC). However, the underlying mechanism of how baicalein targets cellular processes during NSCLC cell invasion and metastasis remains elusive. In this study, we found that non-cytotoxic concentrations of baicalein still retained anti-dissemination activity both in vitro and in vivo. Using a genetic encoding tension probe based on Förster resonance energy transfer (FRET) theory, baicalein was shown to significantly decrease ezrin tension by downregulating cellular ezrin S -nitrosylation (SNO) levels in NSCLC cells in the inflammatory microenvironment. Decreased ezrin tension inhibited the formation of an aggressive phenotype of NSCLC cell and leader cell in collective migration, and subsequently suppressed NSCLC dissemination. Baicalein restrained SNO-mediated ezrin tension by decreasing iNOS expression levels. Overall this study demonstrates the novel mechanism used by baicalein to suppress NSCLC invasion and metastasis from a mechanopharmacology perspective and illustrates a new direction for drug development.
研究领域:
肿瘤学
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药理学
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细胞生物学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2020
·
IF 7.1
Songlin Liu,Hao Ma,Jinyu Zhu,Xiao Xia Han,Bing Zhao
Protein S-nitrosylation is an important post-translational modification, which has important effect on protein functions and is relevant to various diseases. For quantification of the S-nitrosated proteins, it is still challenging to establish a detection method in combination of fast and one-step determination, highly sensitivity and accuracy. Here, we develop a label-free, direct detection method for S-nitrosated proteins based on resonance Raman scattering of ferrous cytochrome c. The laser source of the Raman spectrometer is utilized both for inducing NO leakage and for Raman scattering excitation. The heme ligation alteration and oxidation enable fast quantification of the released NO by the resonance Raman spectroscopy of the cytochrome c. All the results shown in this study demonstrate the great potential of the proposed method for fast, sensitive and reliable assessment of total S-nitrosated proteins in the clinical applications.
研究领域:
生物化学
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医学诊断
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产品关联
DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2021
·
IF 6.953
Pengfei Li,Dawei Jin,Jie Dou,Lijuan Wang,Yanfang Wang,Xingxing Jin,Xiao Han,Inn-Kyu Kang,Jiang Yuan,Jian Shen,Meng Yin
Rapid endothelialization and regulation of smooth muscle cell proliferation are crucial for small-diameter vascular grafts to address poor compliance, thromboembolism, and intimal hyperplasia, and achieve revascularization. As a gaseous signaling molecule, nitric oxide (NO) regulates cardiovascular homeostasis , inhibits blood clotting and intimal hyperplasia, and promotes the growth of endothelial cells. Due to the instability and burst release of small molecular NO donors, a novel biomacromolecular donor has generated increasing interest. In the study, a low toxic NO donor of S-nitrosated keratin (KSNO) was first synthesized and then coelectrospun with poly(ε-caprolactone) to afford NO-releasing small-diameter vascular graft. PCL/KSNO graft was capable to generate NO under the catalysis of ascorbic acid (Asc), so the graft selectively elevated adhesion and growth of human umbilical vein endothelial cells (HUVECs), while inhibited the proliferation of human aortic smooth muscle cells (HASMCs) in the presence of Asc. In addition, the graft displayed significant antibacterial properties and good blood compatibility. Animal experiments showed that the biocomposite graft could inhibit thrombus formation and preserve normal blood flow via single rabbit carotid artery replacement for 1 month. More importantly, a complete endothelium was observed on the lumen surface. Taken together, PCL/KSNO small-diameter vascular graft has potential applications in vascular tissue engineering with rapid endothelialization and vascular remolding.
研究领域:
生物材料学
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心血管医学
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组织工程学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2024
·
IF 12.2
Kunlong Yang,Yue Luo,Tongzheng Sun,Han Qiu,Qingru Geng,Yongxin Li,Man Liu,Nancy P. Keller,Fengqin Song,Jun Tian
Nitric oxide (NO) is a signaling molecule with diverse roles in various organisms. However, its role in the opportunistic pathogen Aspergillus flavus remains unclear. This study investigates the potential of NO, mediated by metabolites from A. oryzae (AO), as an antifungal strategy against A . flavus . We demonstrated that AO metabolites effectively suppressed A . flavus asexual development, a critical stage in its lifecycle. Transcriptomic analysis revealed that AO metabolites induced NO synthesis genes, leading to increased intracellular NO levels. Reducing intracellular NO content rescued A . flavus spores from germination inhibition caused by AO metabolites. Furthermore, exogenous NO treatment and dysfunction of flavohemoglobin Fhb1, a key NO detoxification enzyme, significantly impaired A . flavus asexual development. RNA-sequencing and metabolomic analyses revealed significant metabolic disruptions within tricarboxylic acid (TCA) cycle upon AO treatment. NO treatment significantly reduced mitochondrial membrane potential (Δψm) and ATP generation. Additionally, aberrant metabolic flux within the TCA cycle was observed upon NO treatment. Further analysis revealed that NO induced S-nitrosylation of five key TCA cycle enzymes. Genetic analysis demonstrated that the S-nitrosylated Aconitase Acon and one subunit of succinate dehydrogenase Sdh2 played crucial roles in A . flavus development by regulating ATP production. This study highlights the potential of NO as a novel antifungal strategy to control A . flavus by compromising its mitochondrial function and energy metabolism.
研究领域:
微生物学
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分子生物学
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真菌学
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生物化学
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产品关联
DOI
PubMed
高影响因子
Sensing and Bio-Sensing Research
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2015
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IF 0
Ruirui Wang,Qingnan Kong,Jie Zhou,Lili Zhang,Shuhua Zhu
S -Nitrosylation has recently emerged as an important posttranslational modification of proteins and is becoming an intensive field of research in plants. Protein S -nitrosation, a reversible post-translation modification of cysteine, affects many cell signaling pathways and plays critical roles in redox-sensitive cell signaling. Changes in protein function effectively transmit biological signals and thus provide a framework for elucidating signaling networks. This paper presented a new, universal immunosensor for detection of S -nitrosylated proteins. Electrochemical impedance spectroscopy (EIS) and atomic force microscope (AFM) were used to estimate the formation of self-assembled film. This method was based on the specific binding characteristics of biotin–streptavidin, using Biotin-HPDP labeled protein sulfhydryl group as the substrate to detect proteins. The sensor was used to detect bovine serum albumin (BSA), nitrosylated BSA and denitrosylated BSA. The results showed that 90.61% of nitrosylated BSA were reduced, verifying that protein S -nitrosylation is a reversible and effective post-translation modification. This method was successfully applied to detect S -nitrosylated protein in Feicheng peach. The results showed good repeatability and precision. This method provided a molecular basis for further exploring the mechanism of S -nitrosylation of proteins in plants.
研究领域:
生物化学
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植物科学
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蛋白质组学
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产品关联
DOI
Advanced Science
·
2024
·
IF 14.3
Ping-Fen Yang,Tai-Lei Nie,Xia-Nan Sun,Lan-Xin Xu,Cong Ma,Fang Wang,Li-Hong Long,Jian-Guo Chen
Physical exercise has beneficial effect on anxiety disorders, but the underlying molecular mechanism remains largely unknown. Here, it is demonstrated that physical exercise can downregulate the S-nitrosylation of gephyrin (SNO-gephyrin) in the basolateral amygdala (BLA) to exert anxiolytic effects. It is found that the level of SNO-gephyrin is significantly increased in the BLA of high-anxiety rats and a downregulation of SNO-gephyrin at cysteines 212 and 284 produced anxiolytic effect. Mechanistically, inhibition of SNO-gephyrin by either Cys212 or Cys284 mutations increased the surface expression of GABA A R γ2 and the subsequent GABAergic neurotransmission, exerting anxiolytic effect in male rats. On the other side, overexpression of neuronal nitric oxide synthase in the BLA abolished the anxiolytic-like effects of physical exercise. This study reveals a key role of downregulating SNO-gephyrin in the anxiolytic effects of physical exercise, providing a new explanation for protein post-translational modifications in the brain after exercise.
研究领域:
神经科学
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分子生物学
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行为科学
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产品关联
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2020
·
IF 10.652
Xiuzhen Wan,Shuai Liu,Xuanxuan Xin,Pengfei Li,Jie Dou,Xiao Han,Inn-Kyu Kang,Jiang Yuan,Bo Chi,Jian Shen
Wound repair is a dynamic and complex process that involves various interactions between repair cells, growth factors and the extracellular matrix. Nitric oxide (NO) plays a pivotal role in wound healing, including causing the contraction of wound surfaces, dilating blood vessels, participating in inflammation, and promoting collagen synthesis, angiogenesis, and fibroblast proliferation. Here, we prepared a novel keratin-based nitric oxide donor, called S-nitrosated keratin (KSNO). Then, KSNO was coelectrospun with polyurethane (PU) and gelatin (Gel) to produce PU/Gel/KSNO biocomposite mats. The prepared mats released NO without cytotoxicity, promoted the proliferation of L929 murine fibroblasts and human umbilical vein endothelial cells (HUVECs), and inhibited bacterial growth. These mats could also accelerate the cell adhesion and growth along the random arrangement of electrospun fibers, and simulate the structure of the natural extracellular matrix to provide good scaffolds for cell. The effect of PU/Gel/KSNO on promoting wound healing was investigated on a full-thickness excisional cutaneous wound model in rats. The results demonstrated that these mats accelerated wound healing without inflammatory reaction. In conclusion, PU/Gel/KSNO mats display excellent bioactivity and biocompatibility and have potential in the field of wound repair.
研究领域:
生物医学工程
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材料科学
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组织工程
·
伤口愈合
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产品关联
DOI
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
·
2019
·
IF 3.973
Weilan Li,Jiahui Yong,Yan Xu,Yonglu Wang,Yifan Zhang,Hao Ren,Xueming Li
Photodynamic therapy (PDT) is a prospective approach to cure tumor diseases. However, tumor micro-environment is notably characterized with severe hypoxia and high expression of glutathione (GSH), which seriously limit its clinical application. Here, based on the characteristics of perfluorocarbon (PFC) to dissolve substantial oxygen (O 2 ) and the sensitivity of reductive GSH to S-NO group, we designed GSH depletion and dual-model O 2 supply strategies to promote PDT enhancement. The PFC nanoliposomes ( [email protected] ) and biocompatible NO donor S-nitrosated human serum albumin (HSA-SNO) were combined to synergistically combat the obstacle of tumor micro-environment, reducing GSH concentration and increasing singlet oxygen ( 1 O 2 ) generation. In vitro , after irradiation with NIR laser, the PFC in [email protected] dissolved more O 2 to increase 1 O 2 generation. In addition, with co-delivery of HSA-SNO, it can effectively promote GSH depletion to recover 1 O 2 level and release NO concurrently to inhibit mitochondrial respiration. This combination strategy of [email protected] and HSA-SNO obviously relieved intracellular hypoxia and decreased GSH to increase more toxic 1 O 2 generation for PDT enhancement. The present work will play as an enlightening role in PDT design and clinical application in the near future.
研究领域:
癌症治疗
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生物医学工程
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药学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Acta Biomaterialia
·
2025
·
IF 9.6
Jie Zhang,Yanlong Wang,Yu Sun,Zeyi Zhou,Fengni Liu,Xinyu Zhang,Chuiyu Kong,Jiang Yuan,Meng Yin,Dongjin Wang
Intimal hyperplasia, thrombosis formation, and vascular calcification are the leading causes of failure in small-diameter vascular grafts. In the study, bilayer vascular grafts with separately released nitric oxide (NO) and resveratrol (RESV) were fabricated. S-nitrosated keratin nanoparticles (KNPs) were synthesized and subsequently electrospun with polycaprolactone (PCL) to prolong the NO release for up to 25 d in the presence of ascorbic acid and trypsin, serving as the inner layer of grafts. Additionally, RESV was incorporated into poly(ε-caprolactone-co-L-lactide) (PLCL) fibers to alleviate oxidative stress and inflammation, acting as the outer layer of the grafts. The PCL/KNPs//PLCL/RESV bilayer grafts were capable of promoting endothelial cell proliferation while inhibiting the excessive proliferation of smooth muscle cells. Notably, bilayer grafts could regulate macrophage polarization toward the M2 phenotype. In rat abdominal aorta replacement models, the grafts retained patency for 3 months. These grafts could accelerate endothelialization without apparent intimal hyperplasia, thrombosis, inflammation, and calcification. These bilayer grafts are promising for small-diameter tissue-engineered vascular grafts. Statement of Significance • S-nitrosated keratin nanoparticles (KNPs) were synthesized and subsequently electrospun to prolong the NO release for 25 d. • Resveratrol (RESV) was incorporated into PLCL fibers to alleviate oxidative stress and inflammation. • Bilayer grafts promoted EC proliferation while inhibiting SMC proliferation. • Bilayer grafts accelerated endothelialization without intimal hyperplasia, thrombosis, and calcification in vivo.
研究领域:
生物材料
·
组织工程
·
纳米技术
·
心血管医学
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药学
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生物医学工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Regenerative Biomaterials
·
2022
·
IF 5.763
Dou Jie,Yang Rong,Jin Xingxing,Li Pengfei,Han Xiao,Wang Lijuan,Chi Bo,Shen Jian,Yuan Jiang
Nitric oxide (NO) plays an important role in wound healing, due to its ability to contract wound surfaces, dilate blood vessels, participate in inflammation as well as promote collagen synthesis, angiogenesis and fibroblast proliferation. Herein, keratin was first nitrosated to afford S-nitrosated keratin (KSNO). As a NO donor, KSNO was then co-electrospun with polyurethane (PU). These as-spun PU/KSNO biocomposite mats could release NO sustainably for 72 h, matching the renewal time of the wound dressing. Moreover, these mats exhibited excellent cytocompatibility with good cell adhesion and cell migration. Further, the biocomposite mats exhibited antibacterial properties without inducing severe inflammatory responses. The wound repair in vivo demonstrated that these mats accelerated wound healing by promoting tissue formation, collagen deposition, cell migration, re-epithelialization and angiogenesis. Overall, PU/KSNO mats may be promising candidates for wound dressing.
研究领域:
生物材料
·
伤口愈合
·
组织工程
·
生物医学工程
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产品关联
DOI
PubMed
高影响因子
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