ACTA BIOCHIMICA ET BIOPHYSICA SINICA
·
2018
·Vol. 50
·期 6
·547-554
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IF 2.224
Jia Qian,Lin Yun,Gou Xuejing,He Lei,Shen Dong,Chen Dongni,Xie Wei,Lu Yongjun
The gram-negative bacterium Legionella pneumophila invades human’s lung and causes Legionnaires’ disease. To benefit its survival and replication in cellular milieu, L. pneumophila secrets at least 330 effector proteins into host cells. We found that the effector WipA has the protein tyrosine phosphatase (PTP) activity but does not depend on the classical CX5R motif for activity, suggesting that WipA is an unconventional PTP. Meanwhile, the presence of three other highly conserved motifs typically seen in protein serine/threonine phosphatases and the poor inhibition of WipA activity by okadaic acid led us to propose that WipA is a bacterial protein phosphatase. In addition, the determination of the 2.55-Å crystal structure of WipA revealed that WipA resembles cold-active protein tyrosine phosphatase (CAPTPase), and therefore very likely shares the same catalytic mechanism.
研究领域:
微生物学
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分子生物学
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传染病学
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结构生物学
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DOI
PubMed
高影响因子
Analytical Methods
·
2014
·Vol. 6
·期 15
·6105-6109
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IF 1.938
Fangyuan Wang,Yongxin Li,Wenying Li,Qingfeng Zhang,Jian Chen,Huipeng Zhou,Cong Yu
In the present study, a facile fluorescence turn-on approach for the detection of alkaline phosphatase (ALP) activity has been developed. L-ascorbic acid-2-phosphate (AAP), a substrate of ALP, could be hydrolyzed by ALP to give L-ascorbic acid. L-ascorbic acid then reduced resazurin to resorufin, which resulted in a turn-on fluorescence signal. The fluorescence intensity increase could be directly related to the amount of ALP added to the assay solution. The assay is very sensitive, as 0.12 mU mL−1 ALP could be clearly detected. Sodium orthovanadate (Na3VO4), a well-known ALP inhibitor, was tested, and a clear inhibition effect was observed. The results suggest that our method could be used for ALP activity sensing related various biochemical applications and for the screening of ALP inhibitors.
研究领域:
生物化学
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分析化学
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分子生物学
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DOI
高影响因子
Advanced Science
·
2025
·
IF 14.1
Yating Li,Jason Tan,Yanqin Tu,Jingnan Wu,Zaifang Zhang,Yifeng Wei,Xinan Jiao,Yan Zhou
DNA base modification is a common strategy used by bacteriophages to evade host immune detection. A prominent example is dU-DNA, where thymidine is globally replaced with 2′-deoxyuridine. Despite its widespread occurrence, dU-DNA's biosynthetic pathways and functional roles remain incompletely understood. Here, different enzymes supporting dU-DNA biosynthesis in phage PBS1, Roseophage DSS3_VP1, and Yersiniophage PhiR1-37 are identified and characterized. The dU nucleotide precursor is supplied by phage dCTP deaminases (Dcds). Thymidine nucleotides are degraded by dTMP phosphatases (Dtms) in PBS1 and DSS3_VP1, and by a dTTP pyrophosphatase (Dtt) in PhiR1-37, preventing incorporation into phage DNA. The dU-DNA, isolated from Yersiniophage PhiR1-37 or synthesized by PCR, demonstrates resistance to cleavage by restriction enzymes recognizing thymidine-containing sequences, partial resistance to Lb Cas12a nuclease recognizing a TTTV PAM site, while remaining sensitive to Sp Cas9 nuclease recognizing a NGG PAM site. A phylogenetic analysis of PBS1 dCTP deaminase and closely related T4 phage dCMP deaminase suggest possible evolutionary origins from bacterial dCDP deaminases. Overall, these findings suggest independent acquisition of dU-DNA biosynthetic enzymes and pathways in the diverse phages, and support its protective function against different host-encoded nucleases.
研究领域:
分子生物学
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病毒学
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微生物学
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基因工程
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核酸代谢
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免疫逃避
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进化生物学
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DOI
PubMed
高影响因子
IRISH JOURNAL OF MEDICAL SCIENCE
·
2015
·Vol. 185
·期 3
·581-587
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IF 0.827
Zhao S.,Niu F.,Xu C.-Y.,Liu Y.,Ye L.,Bi G.-B.,Chen L.,Tian G.,Nie T.-H.
Objective To observe the preventive and therapeutic effects of diosgenin on retinoic acid-induced osteoporosis in rats. Methods A total 50 Sprague-Dawley rats were randomly divided into 5 groups: control group, model group (osteoporosis rats), low (10 mg kg −1 ), middle (30 mg kg −1 ), and high-dose diosgenin (90 mg kg −1 )-treated groups. The osteoporosis rats model was induced by retinoic acid. The BMD and physical parameters of femoral including length, wet weight, and dry weight in each group were measured. The hematoxylin–eosin staining was used for bone histomorphology analysis. Besides, the bone calcium (Ca) and phosphorus (P) contents were measured. In order to detect the biochemical index in different treatment groups, the serum tartrate-resistant acid phosphatase (TRAP), alkaline phosphatase (ALP), estradiol, and osteocalcin were compared among different groups. Results The osteoporosis rat model was successfully induced by retinoic acid. Compared with the model group, the lessening of femoral length and weight and the loss of BMD were significantly improved in diosgenin groups. Both contents of Ca and P were much more increased when induced by retinoic acid ( p < 0.05). The estradiol and osteocalcin levels in the middle and high-dose treatment groups were significantly higher than that of the model group, while the ALP and TRAP levels were much lower than the model group ( p < 0.05). Conclusion Diosgenin can prevent the loss of bone in experimental rats. The mechanism may be that it improves the level of estrogenic hormone of estradiol and inhibits the high bone turnover.
研究领域:
药理学
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内分泌学
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骨科
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动物模型研究
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DOI
PubMed
高影响因子
JOURNAL OF NANOBIOTECHNOLOGY
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2019
·Vol. 17
·期 1
·1-11
·
IF 5.345
Hou Zhanwu,Wang Zhen,Liu Run,Li Hua,Zhang Zhengyi,Su Tian,Yang Jeffy,Liu Huadong
Background Gold nanoparticles (AuNPs) have been proposed for many applications in medicine and bioanalysis. For use in all these applications, maintaining the stability of AuNPs in solution by suppressing aggregation is paramount. Herein, the effects of amino acids were investigated in stabilizing AuNPs by rationally designed peptide scaffolds.Results Compared to other tested amino acids, phosphotyrosine (pY) significantly stabilized AuNPs. Our results indicated that pY modified AuNPs presented a high level of stability in various solutions, and had good biocompatibility. When a pY-peptide was used in stabilizing AuNPs, the phosphate group could be removed by phosphatases, which subsequently caused the aggregation and the cargo release of AuNPs. In vitro study showed that AuNPs formed aggregation in a phosphatase concentration depending manner. The aggregation of AuNPs was well correlated with the enzymatic activity (R2 = 0.994). In many types of cancer, a significant increase in phosphatases has been observed. Herein, we demonstrated that cancer cells treated with pY modified AuNPs in conjunction with doxorubicin killed SGC-7901 cells with high efficiency, indicating that the pY peptide stabilized AuNPs could be used as carriers for targeted drug delivery.Conclusion In summary, pY peptides can act to stabilize AuNPs in various solutions. In addition, the aggregation of pY-AuNPs could be tuned by phosphatase. These results provide a basis for pY-AuNPs acting as potential drug carriers and anticancer efficacy.
研究领域:
生物化学
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纳米医学
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肿瘤学
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DOI
PubMed
高影响因子
CHEMOSPHERE
·
2020
·
IF 5.778
Rui Yang,Jinhua Wang,Lusheng Zhu,Jun Wang,Lili Yang,Shushuai Mao,Jeremy L. Conkle,Yangyang Chen,Young Mo Kim
Antibiotics are detected in association with heavy metals in the soil. However, interactions between antibiotics and heavy metals on soil enzyme activity have yet to been studied thoroughly. In this study, soil enzyme activity (urease, sucrase, phosphatase , and Rubisco) were measured after exposure to soils dosed with copper (Cu) and/or enrofloxacin (ENR) over 28 days. Enzyme responses to ENR only treatments varied, but Cu exhibited a strong negative response from all soil enzymes except Rubisco. An interaction between the effects of the two pollutants on soil enzymes was observed in the combined contamination treatments. Greater comprehensive toxicity to soil enzyme activity was observed in combined treatment groups compared to other groups. We anticipate our studies can provide a scientific theoretical basis for the combined pollution of antibiotics and heavy metals in soil.
研究领域:
环境科学
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微生物学
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DOI
PubMed
高影响因子
Talanta Open
·
2024
·
IF 4.1
Chen-Xi Zhao,Xiao-Xia Li,Yang Shu
Abnormal alkaline phosphatase levels have been associated with several diseases, including tumor growth, diabetes, liver dysfunction and so on. Generally, the occurrence and prognosis of these diseases can be reflected by the serum alkaline phosphatase level. In this work, Eu/DBM@SiO 2 @SiO 2 is prepared to protect the internal Eu complex from the external environment by wrapping Eu/DBM inside the silica shell. Then, Eu/DBM@SiO 2 @SiO 2 /MnO 2 , a sensor consists of Eu/DBM@SiO 2 @SiO 2 and MnO 2 , is prepared and applied for alkaline phosphatase detection. MnO 2 is used as the response part of alkaline phosphatase, which can be degraded to Mn 2+ by ascorbic acid. Ascorbic acid is the hydrolysate of alkaline phosphatase. In the process of this reaction, UV–vis absorption of Eu/DBM@SiO 2 @SiO 2 /MnO 2 is reduced and fluorescence is turned on. Alkaline phosphatase concentration is linearly correlated with the logarithmic value of fluorescence enhancement in the range of 10.0–100.0 U/L with linear equation of Lg(F/F 0 )=0.008C-0.030 (R 2 =0.99). For fluorescence method, the LOQ is 10.0 U/L and the LOD is 4.4 U/L. For colorimetric method, the concentration of alkaline phosphatase is linearly correlated with the decrease of absorption intensity in the range of 20.0–90.0 U/L with the linear equation of A/A 0 =-0.012C+1.148 (R 2 =0.99). The LOQ is 20.0 U/L and the LOD is 7.2 U/L. The detection system has good selectivity and can detect alkaline phosphatase in human serum samples, the accuracy of the detection system is verified by the experiment of spike-in experiment. The RSD of alkaline phosphatase concentration in serum samples is in the range of 1.4–8.4 %. This method has potential application in the detection of ALP activity in biological samples.
研究领域:
生物化学
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医学诊断
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DOI
高影响因子
CHEMICAL & PHARMACEUTICAL BULLETIN
·
2020
·Vol. 68
·期 7
·628-634
·
IF 1.416
Jianfeng Pan,Chenfang Miao,Yuanting Chen,Jiahui Ye,Zhenzhen Wang,Wendi Han,Zhengjun Huang,Yanjie Zheng,Shaohuang Weng
Facile and effective detection of dopamine (DA) plays a significant role in current clinical applications. Substantially, special optical nanomaterials are important for fabricating easy-to-control, cheap, selective, and portable fluorescence DA sensors with superior performance. Herein, carbon dots (CDs) prepared from melting method were applied as signal to establish a simple but effective fluorescence strategy for DA determination based on the enzymatic activity of acid phosphatase (ACP), which induces DA to form polydopamine (pDA). The formed pDA caused by the enzymatic oxidization of ACP toward DA can interact with CDs through the inner filter effect. Such behavior effectively quenched the CDs’ fluorescence. The degree of fluorescence quenching of CDs was positively correlated with the DA content. Under the optimized reaction conditions, the proposed fluorescence method exhibited a comparable analytical performance with other DA sensors with good selectivity. Furthermore, this method has been successfully applied to detect DA in DA hydrochloride injection and human serum samples. It shows that this method features potential practical application value and is expected to be used in clinical research.
研究领域:
生物化学
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医学检测
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纳米材料科学
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DOI
PubMed
高影响因子
Journal of Materials Chemistry B
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2025
·
IF 5.7
Yingxian Shi,Ruirui Dong,Huan Huang,Gaoying Wang,Jianyi Gao,Kai Wang,Ting Zhang
Ovarian cancer (OC) remains a prevalent gynecological malignancy, posing significant threats to women's health globally. Emerging evidence underscores the critical involvement of alkaline phosphatase (ALP) in OC pathogenesis, prompting the development of targeted diagnostic tools. Herein, we report the successful application of a novel fluorescent probe, HD-ALP, for fluorescent imaging of ALP in ovarian cancer cells and preclinical models. After being triggered by ALP, HD-ALP responded in the fluorescent signal reporting manner. More importantly, the fluorescence signal showed a reliable linear correlation with ALP levels in the range of 0-10000 U/L. The HD-ALP probe demonstrated high sensitivity, selectivity, and stability under the necessary biological conditions. The intracellular imaging results indicated that HD-ALP enabled the visualization of ALP levels within ovarian cancer cells. The in vivo experiments demonstrated that HD-ALP could accurately differentiate between healthy and cancerous models (subcutaneous transplantation tumor model of ovarian cancer, orthotopic ovarian cancer model, primary ovarian cancer model). Notably, In cellular and in vivo imaging, HD-ALP could effectively reflect the decrease in fluorescence signal after the addition of ALP inhibitors. By integrating ALP detection, which utilizes the HD-ALP fluorescence mode, with traditional methods like immunohistochemistry staining and serum biomarker detection, potential strategies for more accurate diagnosis of OC may be developed for preclinical trials in the future. According to the research, this is also one of the few developments of fluorescent detection tools for measuring ALP levels in ovarian cancer models.
研究领域:
肿瘤学
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医学影像学
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生物化学
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DOI
PubMed
高影响因子
ENVIRONMENTAL TOXICOLOGY AND CHEMISTRY
·
2020
·Vol. 40
·期 2
·342-351
·
IF 3.152
Wenjing Wang,Ming Jiang,Yanqing Sheng
Glyphosate is a commonly used herbicide known for its high performance in killing certain plants and grasses; however, its use is regulated due to its harmful effects on the aquatic environment. The present study investigated and compared the toxic mechanisms of glyphosate on Microcystis aeruginosa (a toxin-producing cyanobacterium) under 2 conditions: 0‰ saline media (experiment I) and 2.5‰ saline media (experiment II). The results indicated that an appropriate concentration of glyphosate provided a phosphate source for M. aeruginosa , resulting in an increased specific growth rate in both experimental groups compared with the controls. Glyphosate-enhanced alkaline phosphatase (ALP) activity increased by up to 1.37-fold in experiment I and 1.68-fold in experiment II. Moreover, the activities of superoxide dismutase (SOD) and catalase (CAT) decreased at glyphosate concentrations below 1.2 mg L –1 but increased at concentrations greater than 1.2 mg L –1 in experiment I, whereas SOD and CAT activities decreased in experiment II and declined by 64 and 49% in the 30 mg L −1 treatments. Furthermore, the transcript abundances of the pyruvate carboxylase ( pcB ), microcystin synthetase B ( mcyB ) , and paired-like homeobox ( phoX ) genes were up-regulated by up to 6.92-, 3.63-, and 2.27-fold in experiment I and 6.74-, 6.55-, and 4.86-fold in experiment II after 96 h of incubation. The addition of glyphosate stimulated the production of dissolved organic matter including tryptophan-like substances, fulvic acid-like substances, (marine) humic acid-like substances, and microcystin–leucine-arginine in the culture. In conclusion, glyphosate stimulates the proliferation of M. aeruginosa and enhances the release of dissolved organic matter in saltwater ecosystems. Environ Toxicol Chem 2021;40:342–351. © 2020 SETAC
研究领域:
环境毒理学
·
水生生态学
·
分子生物学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2019
·
IF 6.393
Yuyuan Chen,Zhenzhen Wang,Xiaoli Hao,Fenglan Li,Yanjie Zheng,Jianzhong Zhang,Xinhua Lin,Shaohuang Weng
Detection of acid phosphatase (ACP), a universal and important enzyme, is crucial in monitoring several diseases. However, the selectivity of ACP detection under neutral conditions is poor. Herein, ACP was found to catalyze dopamine as a new substrate to polydopamine in acidic or neutral conditions, whereas alkaline phosphatase (ALP) failed to serve such function. The catalytic mechanism underlying the formation of polydopamine was investigated. A simple assay for ACP was developed using fluorescent carbon dots as a signal indicator. In the presence of ACP, dopamine polymerization was conducted in situ and carbon dots were quenched through the inner filter effect. This fluorescent bioassay illustrated a good analytical performance with a wide linear range and a detection limit of 0.45 U/L. In particular, ACP showed high selectivity from the interference of ALP under neutral conditions. Furthermore, excellent test recovery in human serum samples demonstrated the predictable applications of this method in clinical and biomedical areas.
研究领域:
生物化学
·
医学检测
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分析化学
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产品关联
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2024
·
IF 8.3
Jieqiong Chen,Qing Zhao,Jiajing Tang,Xiaoyu Lei,Jinzheng Zhang,Yuping Li,Jidong Li,Yubao Li,Yi Zuo
Inspired by the crucial role of matrix vesicles (MVs), a series of biomimetic vesicles (BVs) fabricated by calcium glycerophosphate (CaGP) modified polyurethane were designed to mediate the mineralization through in situ enzyme activation for bone therapy. In this study, alkaline phosphatase (ALP) was harbored in the porous BVs by adsorption (Ad-BVs) or entrapment (En-BVs). High encapsulation of ALP on En-BVs was effectively self-activating by calcium ions of CaGP-modified PU that specifically hydrolyzed the organophosphorus (CaGP) to inorganic phosphate, thus promoting the formation of the highly oriented bone-like apatite in vitro. Enzyme-catalyzed kinetics confirms the regulation of apatite crystallization by the synergistic action of self-activated ALP and the confined microcompartments of BVs. This leads to a supersaturated microenvironment, with the En-BVs group exhibiting inorganic phosphate (Pi) levels 4.19 times higher and Ca2+ levels 3.67 times higher than those of simulated body fluid (SBF). Of note, the En-BVs group exhibited excellent osteo-inducing differentiation of BMSCs in vitro and the highest maturity with reduced bone loss in rat femoral defect in vivo. This innovative strategy of biomimetic vesicles is expected to provide valuable insights into the enzyme-activated field of bone therapy.
研究领域:
生物材料学
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骨再生医学
·
组织工程
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）