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1.
Alan H. Johnson John R. Baker 《Biochimica et Biophysica Acta (BBA)/General Subjects》1973,320(2):341-351
A sulphotransferase preparation from hen's uterus catalysed the transfer of sulphate from adenosine 3′-phosphate 5′-sulphatophosphate to N-desulphated heparan sulphate, heparan sulphate, N-desulphated heparin and dermatan sulphate. Heparin, chondroitin sulphate and hyaluronic acid were inactive as substrates for the enzyme. N-desulphated heparin was a much poorer substrate for the enzyme than N-desulphated heparan sulphate suggesting that properties of the substrate other than available glucosaminyl residues influenced enzyme activity. N-acetylation of N-desulphated heparin and N-desulphated heparan sulphate reduced their sulphate acceptor properties so it was unlikely that the N-acetyl groups of heparan sulphate facilitated its sulphatiion. Direct evidence for the transfer of [35S]sulphate to amino groups of N-desulphated haparan sulphate was obtained by subsequent isolation of glucosamine from heparan [35S]sulphate product. This was made possible through the use of a flavobacterial enzyme preparation which contained “heparitinase” activity but had been essentially freed of sulphatases. Attempts to transfer [35S]sulphate to glucosamine or N-acetylglucosamine were unsuccessfull. 相似文献
2.
Yuan Lu Yan Dong Yan Zhang Di Shen Xiyao Wang Ruxiu Ge Meihua Zhang Yu Xia Xietong Wang 《Journal of cellular and molecular medicine》2020,24(12):6690-6703
Despite the widespread use of antiplatelets and anticoagulants, women with antiphospholipid syndrome (APS) may face pregnancy complications associated with placental dysplasia. Neutrophil extracellular traps (NETs) are involved in the pathogenesis of many autoimmune diseases, including vascular APS; however, their role in obstetric APS is unclear. Herein, we investigated the role of NETs by quantifying cell‐free DNA and NET marker levels. Live‐cell imaging was used to visualize NET formation, and MAPK signalling pathway proteins were analysed. Cell migration, invasion and tube formation assays were performed to observe the effects of NETs on trophoblasts and human umbilical vein endothelial cells (HUVECs). The concentrations of cell‐free DNA and NETs in sera of pregnant patients with APS were elevated compared with that of healthy controls (HCs) matched to gestational week. APS neutrophils were predisposed to spontaneous NET release and IgG purified from the patients (APS‐IgG) induced neutrophils from HCs to release NETs. Additionally, APS‐IgG NET induction was abolished with inhibitors of reactive oxygen species, AKT, p38 MAPK and ERK1/2. Moreover, NETs were detrimental to trophoblasts and HUVECs. In summary, APS‐IgG‐induced NET formation deserves further investigation as a potential novel therapeutic target in obstetrical APS. 相似文献
3.
Elias Silva dos Santos Leila Cristiane Virgens de Souza Patrícia Nascimento de Assis Elias Ramos-de-Souza 《Journal of biomolecular structure & dynamics》2013,31(6):1176-1184
We have used docking techniques in order to propose potential inhibitors to the enzymes adenosine phosphosulfate reductase and adenosine triphosphate sulfurylase that are responsible, among other deleterious effects, for causing souring of oil and gas reservoirs. Three candidates selected through molecular docking revealed new and improved polar and hydrophobic interactions with the above-mentioned enzymes. Microbiological laboratory assays performed subsequently corroborated the results of computer modelling that the three compounds can efficiently control the biogenic sulfide production. 相似文献
4.
Preparation and in vitro evaluation of xanthan gum facilitated superabsorbent polymeric microspheres
Shiv Sankar Bhattacharya Farhan Mazahir Subham Banerjee Anurag Verma Amitava Ghosh 《Carbohydrate polymers》2013
Interpenetrating polymer network (IPN) hydrogel microspheres of xanthan gum (XG) based superabsorbent polymer (SAP) and poly(vinyl alcohol) (PVA) were prepared by water-in-oil (w/o) emulsion crosslinking method for sustained release of ciprofloxacin hydrochloride (CIPRO). The microspheres were prepared with various ratios of hydrolyzed SAP to PVA and extent of crosslinking density. The prepared microspheres with loose and rigid surfaces were evidenced by scanning electron microscope (SEM). Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analysis confirmed the IPN formation. Differential scanning calorimetry (DSC) study was performed to understand the dispersion nature of drug after encapsulation. The in vitro drug release study was extensively evaluated depending on the process variables in both acidic and alkaline media. All the formulations exhibited satisfactory physicochemical and in vitro release characteristics. Release data indicated a non-Fickian trend of drug release from the formulations. Based on the results, this study suggest that CIPRO loaded IPN microspheres were suitable for sustained release application. 相似文献
5.
Pyruvate carboxylase (PC) is a biotin-dependent enzyme that catalyzes the MgATP- and bicarbonate-dependent carboxylation of pyruvate to oxaloacetate, an important anaplerotic reaction in central metabolism. The carboxyltransferase (CT) domain of PC catalyzes the transfer of a carboxyl group from carboxybiotin to the accepting substrate, pyruvate. It has been hypothesized that the reactive enolpyruvate intermediate is stabilized through a bidentate interaction with the metal ion in the CT domain active site. Whereas bidentate ligands are commonly observed in enzymes catalyzing reactions proceeding through an enolpyruvate intermediate, no bidentate interaction has yet been observed in the CT domain of PC. Here, we report three X-ray crystal structures of the Rhizobium etli PC CT domain with the bound inhibitors oxalate, 3-hydroxypyruvate, and 3-bromopyruvate. Oxalate, a stereoelectronic mimic of the enolpyruvate intermediate, does not interact directly with the metal ion. Instead, oxalate is buried in a pocket formed by several positively charged amino acid residues and the metal ion. Furthermore, both 3-hydroxypyruvate and 3-bromopyruvate, analogs of the reaction product oxaloacetate, bind in an identical manner to oxalate suggesting that the substrate maintains its orientation in the active site throughout catalysis. Together, these structures indicate that the substrates, products and intermediates in the PC-catalyzed reaction are not oriented in the active site as previously assumed. The absence of a bidentate interaction with the active site metal appears to be a unique mechanistic feature among the small group of biotin-dependent enzymes that act on α-keto acid substrates. 相似文献
6.
Enhancement of the innate immune response of bladder epithelial cells by Astragalus polysaccharides through upregulation of TLR4 expression 总被引:1,自引:0,他引:1
Xiaolin Yin Lei Chen Jianling Yang Zhiyan Yao Lin Wei Mingyuan Li 《Biochemical and biophysical research communications》2010,397(2):232-238
The innate host defenses at mucosal surfaces are critical in the early stages of urinary tract bacterial infection. Recent studies have shown that uroepithelial cells aid innate immune cells in fighting off infection, although the exact mechanism by which the uroepithilium participates in immunity remains unclear. TLR4 has been implicated to possess antimicrobial activities specific for bladder epithelial cells (BECs). TLR4 promotes secretion of IL-6 and IL-8, mediates inhibition of bladder epithelial cell (BEC) bacterial invasion, and mediates expulsion of uropathogenic Escherichia coli from BECs. In this study, cultured 5637 cells and Balb/C mice were treated with Astragalus polysaccharides (APS) against invading E. coli. To determine the contribution of TLR4 upregulation to immune response, TLR4 expression and bacterial colony numbers were monitored. After 24 h incubation, only 5637 cells treated with 500 μg/ml APS expressed higher levels of TLR4 compared with the untreated group. However, after 48 h, all 5637 cells treated by APS showed higher levels of TLR4 expression than the control cells. The TLR4 expression in the bladder and macrophages mice that received APS was higher than that in the controls. Bacterial colonization in 5637 cells and the bladders of mice treated with APS was significantly reduced compared with the controls. These results demonstrate that at certain concentrations, APS can induce increased TLR4 expression in vivo and in vitro. Further, TLR4 expression upregulation can enhance innate immunity during mucosal bacterial infection. The findings establish the use of APS to modulate the innate immune response of the urinary tract through TLR4 expression regulation as an alternative option for UTI treatment. 相似文献
7.
Sekulic N Dietrich K Paarmann I Ort S Konrad M Lavie A 《Journal of molecular biology》2007,367(2):488-500
Bifunctional human PAPS synthetase (PAPSS) catalyzes, in a two-step process, the formation of the activated sulfate carrier 3'-phosphoadenosine 5'-phosphosulfate (PAPS). The first reaction involves the formation of the 5'-adenosine phosphosulfate (APS) intermediate from ATP and inorganic sulfate. APS is then further phosphorylated on its 3'-hydroxyl group by an additional ATP molecule to generate PAPS. The former reaction is catalyzed by the ATP-sulfurylase domain and the latter by the APS-kinase domain. Here, we report the structure of the APS-kinase domain of PAPSS isoform 1 (PAPSS1) representing the Michaelis complex with the products ADP-Mg and PAPS. This structure provides a rare glimpse of the active conformation of an enzyme catalyzing phosphoryl transfer without resorting to substrate analogs, inactivating mutations, or catalytically non-competent conditions. Our structure shows the interactions involved in the binding of the magnesium ion and PAPS, thereby revealing residues critical for catalysis. The essential magnesium ion is observed bridging the phosphate groups of the products. This function of the metal ion is made possible by the DGDN-loop changing its conformation from that previously reported, and identifies these loop residues unambiguously as a Walker B motif. Furthermore, the second aspartate residue of this motif is the likely candidate for initiating nucleophilic attack on the ATP gamma-phosphate group by abstracting the proton from the 3'-hydroxyl group of the substrate APS. We report the structure of the APS-kinase domain of human PAPSS1 in complex with two APS molecules, demonstrating the ability of the ATP/ADP-binding site to bind APS. Both structures reveal extended N termini that approach the active site of the neighboring monomer. Together, these results significantly increase our understandings of how catalysis is achieved by APS-kinase. 相似文献
8.
黄芪多糖对齐口裂腹鱼生长、体组成和免疫指标的影响 总被引:7,自引:0,他引:7
试验研究黄芪多糖对齐口裂腹鱼生长性能、体组成及免疫指标的影响。以450尾健康的齐口裂腹鱼[体重(6.98±0.43)g;体长(9.11±0.25)cm]为试验对象,随机分为5组(C1、C2、C3、C4、C5),每组3个重复,每重复30尾试验鱼。C1、C2、C3、C4、C5组分别投喂在等氮等能(蛋白质含量38.29%,能量15.73 mJ/kg)的基础料中分别添加0、0.02、0.04、0.06、0.08%的黄芪多糖制成5种试验饲料,养殖齐口裂腹鱼50d。结果表明:饲料中未添加黄芪多糖组的增重率(WGR)、特定生长率(SGR)、饲料蛋白效率(PER)均显著低于黄芪多糖添加组(P<0.05),而饵料系数(FCR)则显著高于黄芪多糖添加组(P<0.05)。当黄芪多糖添加水平为0.04%时,试验鱼的WGR、SGR、PER均达到最大(分别为110.31%、1.86%/d和182.07%),FCR最低(1.44),与其他各组差异显著(P<0.05);以WGR、SGR、PER、FCR为指标,利用直线和抛物线回归分析表明,齐口裂腹鱼生长性能最佳时黄芪多糖添加水平为0.045%—0.074%;对齐口裂腹鱼机体组成分析表明,黄芪多糖对鱼体粗灰分和水分影响不显著(P>0.05),黄芪多糖添加水平为0.06%时机体粗蛋白最高,但与黄芪多糖添加水平为0.04%时无明显差异(P>0.05);粗脂肪含量在黄芪多糖添加水平为0.04%时最高,与其他各组差异显著(P<0.05);未添加黄芪多糖组血清免疫酶活性显著低于黄芪多糖添加组,齐口裂腹鱼血清免疫酶活性在一定范围内随黄芪多糖的增加而增强。黄芪多糖添加水平为0.04%时,碱性磷酸酶(ALP)活性最高;酸性磷酸酶(ACP)活性在黄芪多糖添加水平0.06%时最大;黄芪多糖添加水平应在0.06%—0.08%时,溶菌酶(LSZ)、超氧化歧化酶(SOD活性趋于稳定。这说明黄芪多对齐口裂腹鱼的生长和免疫力有明显的促进作用。综合考虑齐口裂腹鱼生长性能和免疫能力最佳时黄芪多糖添加水平为0.04%—0.074%。 相似文献
9.
Miyuki Bohgaki Masaki Matsumoto Tatsuya Atsumi Takeshi Kondo Shinsuke Yasuda Tetsuya Horita Keiichi I. Nakayama Fumihiko Okumura Shigetsugu Hatakeyama Takao Koike 《Journal of cellular and molecular medicine》2011,15(1):141-151
Antiphospholipid syndrome (APS) is characterized by thrombosis and the presence of antiphospholipid antibodies (aPL) that directly recognizes plasma β2‐glycoprotein I (β2GPI). Tissue factor (TF), the major initiator of the extrinsic coagulation system, is induced on monocytes by aPL in vitro, explaining in part the pathophysiology in APS. We previously reported that the mitogen‐activated protein kinase (MAPK) pathway plays an important role in aPL‐induced TF expression on monocytes. In this study, we identified plasma gelsolin as a protein associated with β2GPI by using immunoaffinity chromatography and mass spectrometric analysis. An in vivo binding assay showed that endogenous β2GPI interacts with plasma gelsolin, which binds to integrin a5β1 through fibronectin. The tethering of β2GPI to monoclonal anti‐β2GPI autoantibody on the cell surface was enhanced in the presence of plasma gelsolin. Immunoblot analysis demonstrated that p38 MAPK protein was phosphorylated by monoclonal anti‐β2GPI antibody treatment, and its phosphorylation was attenuated in the presence of anti‐integrin a5β1 antibody. Furthermore, focal adhesion kinase, a downstream molecule of the fibronectin‐integrin signalling pathway, was phosphorylated by anti‐β2GPI antibody treatment. These results indicate that molecules including gelsolin and integrin are involved in the anti‐β2GPI antibody‐induced MAPK pathway on monocytes and that integrin is a possible therapeutic target to modify a prothrombotic state in patients with APS. 相似文献
10.
Selenium uptake,translocation, assimilation and metabolic fate in plants 总被引:24,自引:0,他引:24
The chemical and physical resemblance between selenium (Se) and sulfur (S) establishes that both these elements share common
metabolic pathways in plants. The presence of isologous Se and S compounds indicates that these elements compete in biochemical
processes that affect uptake, translocation and assimilation throughout plant development. Yet, minor but crucial differences
in reactivity and other metabolic interactions infer that some biochemical processes involving Se may be excluded from those
relating to S. This review examines the current understanding of physiological and biochemical relationships between S and
Se metabolism by highlighting their similarities and differences in relation to uptake, transport and assimilation pathways
as observed in Se hyperaccumulator and non-accumulator plant species. The exploitation of genetic resources used in bioengineering
strategies of plants is illuminating the function of sulfate transporters and key enzymes of the S assimilatory pathway in
relation to Se accumulation and final metabolic fate. These strategies are providing the basic framework by which to resolve
questions relating to the essentiality of Se in plants and the mechanisms utilized by Se hyperaccumulators to circumvent toxicity.
In addition, such approaches may assist in the future application of genetically engineered Se accumulating plants for environmental
renewal and human health objectives. 相似文献