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A preparation strategy for protein-oriented immobilized silica magnetic beads with Spy chemistry for ligand fishing 认领 引用 被引量：1
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作者 Yu Yi Jianming Hu +5 位作者 Shenwei Ding Jianfeng Mei Xudong Wang Yanlu Zhang Jianshu Chen Guoqing Ying 《Journal of Pharmaceutical Analysis》 SCIE CAS CSCD 2022年第3期415-423,共9页
Due to the complexity of bioactive ingredients in biological samples,the screening of target proteins is a complex process.Herein,a feasible strategy for directing protein immobilization on silica magnetic beads for l... Due to the complexity of bioactive ingredients in biological samples,the screening of target proteins is a complex process.Herein,a feasible strategy for directing protein immobilization on silica magnetic beads for ligand fishing based on SpyTag/SpyCatcher(ST/SC)-mediated anchoring is presented.Carboxyl functional groups on the surface of silica-coated magnetic beads(SMBs)were coupled with SC using the 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysulfosuccinimide method,named SC-SMBs.The green fluorescent protein(GFP),as the capturing protein model,was ST-labeled and anchored at a specific orientation onto the surface of SC-SMBs directly from relevant cell lysates via ST/SC self-ligation.The characteristics of the SC-SMBs were studied via electron microscopy,energy dispersive spectroscopy,and Fourier transform infrared spectroscopy.The spontaneity and site-specificity of this unique reaction were confirmed via electrophoresis and fluorescence analyses.Although the alkaline stability of ST-GFP-ligated SC-SMBs was not ideal,the formed isopeptide bond was unbreakable under acidic conditions(0.05 M glycine-HCl buffer,pH 1e6)for 2 h,under 20%ethanol solution within 7 days,and at most temperatures.We,therefore,present a simple and universal strategy for the preparation of diverse protein-functionalized SMBs for ligand fishing,prompting its usage on drug screening and target finding. 展开更多
关键词 EDC/Sulfo-NHS SpyTag/SpyCatcher Green fluorescent protein Silica magnetic beads Oriented immobilization Drug screening
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Production of biliverdin by biotransformation of exogenous heme using recombinant Pichia pastoris cells 认领 引用 被引量：1
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作者 Jianfeng Mei Yanchao Han +3 位作者 Shihang Zhuang Zhikai Yang Yu Yi Guoqing Ying 《Bioresources and Bioprocessing》 SCIE EI 2024年第1期263-272,共10页
Biliverdin,a bile pigment hydrolyzed from heme by heme oxygenase(HO),serves multiple functions in the human body,including antioxidant,anti-inflammatory,and immune response inhibitory activities.Biliverdin has great p... Biliverdin,a bile pigment hydrolyzed from heme by heme oxygenase(HO),serves multiple functions in the human body,including antioxidant,anti-inflammatory,and immune response inhibitory activities.Biliverdin has great potential as a clinical drug;however,no economic and efficient production method is available currently.Therefore,the production of biliverdin by the biotransformation of exogenous heme using recombinant HO-expressing yeast cells was studied in this research.First,the heme oxygenase-1 gene(HO1)encoding the inducible plastidic isozyme from Arabidopsis thaliana,with the plastid transport peptide sequence removed,was recombined into Pichia pasto-ris GS115 cells.This resulted in the construction of a recombinant P.pastoris GS115-HO1 strain that expressed active HO1 in the cytoplasm.After that,the concentration of the inducer methanol,the induction culture time,the pH of the medium,and the concentration of sorbitol supplied in the medium were optimized,resulting in a significant improvement in the yield of HO1.Subsequently,the whole cells of GS115-HO1 were employed as catalysts to convert heme chloride(hemin)into biliverdin.The results showed that the yield of biliverdin was 132 mg/L when hemin was added to the culture of GS115-HO1 and incubated for 4 h at 30°C.The findings of this study have laid a good foundation for future applications of this method for the economical production of biliverdin. 展开更多
关键词 Biliverdin Biotransformation Heme Heme oxygenase-1 Recombinant Pichia pastoris
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Dual roles and therapeutic targeting of tumor-associated macrophages in tumor microenvironments 认领 引用 被引量：31
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作者 Jiasheng Xu Lei Ding +6 位作者 Jianfeng Mei Yeting Hu Xiangxing Kong Siqi Dai Tongtong Bu Qian Xiao Kefeng Ding 《Signal Transduction and Targeted Therapy》 SCIE CSCD 2025年第9期5053-5081,共29页
Tumor-associated macrophages(TAMs),derived from circulating monocytes recruited to tumor sites via chemotactic signals such as C-C motif ligand 2(CCL2)and colony-stimulating factor-1(CSF-1),are pivotal components of t... Tumor-associated macrophages(TAMs),derived from circulating monocytes recruited to tumor sites via chemotactic signals such as C-C motif ligand 2(CCL2)and colony-stimulating factor-1(CSF-1),are pivotal components of the tumor microenvironment(TME).Functionally polarized into distinct subtypes,TAMs play dual roles:proinflammatory M1-type TAMs enhance antitumor immunity through the secretion of cytokines such as interleukin-12(IL-12)and tumor necrosis factor alpha(TNF-α)and direct tumor cell cytotoxicity,whereas M2-type TAMs promote tumor progression by facilitating angiogenesis,metastasis,and immunosuppression.This polarization is dynamically regulated by different cytokines,various signaling pathways,and metabolic cues within the TME.Spatial distribution analyses revealed that M2-like TAMs predominantly infiltrate hypoxic and stromal regions,where they secrete factors such as vascular endothelial growth factor(VEGF),transforming growth factor beta(TGF-β),and matrix metalloproteinases(MMPs)to remodel the extracellular matrix and suppress immune responses via programmed death-ligand 1(PD-L1)and arginase-1 upregulation.Crucially,TAMs interact extensively with immune cells;M2-TAMs secrete interleukin-10(IL-10)and TGF-βto inhibit cytotoxic T lymphocytes while expanding regulatory T(Treg)cells and impairing natural killer(NK)cell function via altered antigen presentation.Conversely,M1-TAMs synergize with dendritic cells to enhance T-cell priming.Therapeutically,targeting TAMs offers promising strategies,including colony-stimulating factor-1 receptor(CSF-1R)inhibitors,CCL2 antagonists,and nanoparticle-mediated repolarization of M2-TAMs toward the M1 phenotype.Emerging genetic approaches,such as clustered regularly interspaced short palindromic repeat-CRISPR-associated protein 9(CRISPR-Cas9)editing,aim to disrupt protumorigenic pathways in TAMs.Additionally,TAM-related biomarkers(e.g.,CD206 and CD163)are being evaluated for their prognostic and predictive utility in immunotherapies.Despite progress,challenges persist owing to TAM plasticity and TME heterogeneity across cancers.This review synthesizes TAM biology,immune crosstalk,and therapeutic advancements,providing a foundation for novel oncology strategies aimed at reprogramming TAMs to overcome treatment resistance and improve clinical outcomes. 展开更多
关键词 enhance antitumor immunity therapeutic targeting chemotactic signals circulating monocytes direct tumor cell cytotoxicitywherea tumor microenvironment tme functionally dual roles tumor microenvironment
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