Transfer Of Sulfate Research Articles

Biocatalytic sulfation of aromatic and aliphatic alcohols catalyzed by arylsulfate sulfotransferases

Many relevant metabolites, as well as chemical commodities, contain at least one sulfate ester group. Consequently, biocatalytic strategies to attach sulfate to a molecule under mild conditions are of high interest. In order to expand the enzymatic toolbox available, five new arylsulfate sulfotransferases (ASSTs) were identified in this study. Overexpression in Escherichia coli and enzyme purification resulted in soluble proteins which catalyzed the sulfate transfer to an acceptor substrate using p-nitrophenyl sulfate (pNPS) as sulfate donor. Optimal reaction conditions were established with respect to temperature and pH, as well as their tolerance to organic co-solvents and melting temperature. Additionally, the kinetic parameters (Vmax, KM, and kcat) were determined. The substrate scope for the acceptor showed that a structurally diverse spectrum of alcohols is accepted. The substrates included phenolic alcohols with one, two, and three hydroxy groups, linear and cyclic aliphatic alcohols, and amines. The phenolic substrates were accepted reaching activities of up to 154 U/mg purified enzyme. Additionally, also the aliphatic alcohols (both linear and cyclic) were accepted at reduced activity, showing that these enzymes are not limited to phenolic alcohols. Moreover, catalytic activity was detected when using aniline as an acceptor substrate implying their ability to sulfate also amino groups. Finally, the consecutive sulfation of di- and trihydroxy compounds was observed, resulting in the detection of the corresponding disulfated molecules.Key points• Five novel arylsulfate sulfotransferases were identified and characterized.• Accepted substrates included aromatic and aliphatic alcohols, as well as aniline.• Disulfation of di- and trihydroxy aromatic compounds was studied and confirmed.

Virus communities rather than bacterial communities contribute more on nutrient pool in polluted aquatic environment

The degradation of animal carcasses can lead to rapid waste release (e.g., pathogenic bacteria, viruses, prions, or parasites) and also result in nutrient accumulation in the surrounding environment. However, how viral profile responds and influences nutrient pool (carbon (C), nitrogen (N), phosphorus (P) and sulfur (S)) in polluted water caused by animal carcass decomposition had not been explored. Here, we combined metagenomic analysis, 16S rRNA gene sequencing and water physicochemical assessment to explore the response of viral communities under different temperatures (23 °C, 26 °C, 29 °C, 32 °C, and 35 °C) in water polluted by cadaver, as well as compare the contribution of viral/bacterial communities on water nutrient pool. We found that a total of 15,240 viral species were classified and mainly consisted of Siphoviridae. Both temperature and carrion reduced the viral diversity and abundance. Only a small portion of the viruses (∼8.8 %) had significant negative correlations with temperature, while most were not sensitive. Our results revealed that the viruses had lager contribution on nutrient pool than bacteria. Besides, viral-related functional genes involved in C, N, P and S cycling. These functional genes declined during carcass decomposition and covered part of the central nutrient cycle metabolism (including carbon sugar transformation, denitrification, P mineralization and extracelluar sulfate transfer, etc.). Our result implies that human regulation of virus communities may be more important than bacterial communities in regulating and managing polluted water quality and nutrition

The sulfate transfer characteristics in recycled aggregate concrete incorporated with fly ash under percolation theory

Recycled aggregate concrete (RAC) has multiple interface transition zones (ITZs) due to its complex composition, providing convenient ion transfer channels. Therefore, it is significant to explore the changes of the internal pore structure in RAC under ion erosion conditions for quantifying and predicting ion transfer mechanisms. This paper innovatively utilizes the seepage effect for investigating the evolution of pore structures in RAC under sulfate attack. We conduct a systematic experimental study on the transport of sulfate ions in RAC with different fly ash (FA) substitution rates under dry-wet cycles. Combining backscattered electron imaging (BSE), permeation and fractal characteristics, this paper deeply explore the relationship between pore structure feature, sulfate percolation mechanism and the macroscopic properties of RAC incorporated with different amount of FA.

Pancancer analysis of the correlations of HS6ST2 with prognosis, tumor immunity, and drug resistance

HS6ST2 has ability to encodes a member of the heparan sulfate (HS) sulfotransferase gene family, which catalyze the transfer of sulfate to HS and a crucial regulator of cell growth, differentiation, adhesion, and migration. Although mounting evidence supports a vital role for HS6ST2 in tumorigenesis of some cancers, no pan-cancer analysis of HS6ST2 has been reported. Therefore, we aimed to explore the prognostic value of HS6ST2 in 33 cancer types and investigate its potential immune function. Based on data from The Cancer Genome Atlas, Cancer Cell Lines Encyclopedia, Genotype Tissue Expression, and GSCA, we used a range of bioinformatics approaches to explore the potential carcinogenic role of HS6ST2, analysis of HS6ST2 and prognosis, DNA methylation, RNA methylation, microsatellite instability (MSI), tumor mutation burden (TMB), and immune cell infiltration in different tumors. The results show that HS6ST2 was highly expressed in most cancers but lower in Breast invasive carcinoma, Kidney Chromophobe, Kidney renal clear cell carcinoma, Kidney renal papillary cell carcinoma, and Uterine Corpus Endometrial Carcinoma. Moreover, HS6ST2 is positively or negatively associated with prognosis in different cancers. HS6ST2 expression was not only associated with MSI in 5 cancer types and associated with TMB in 10 cancer types, and it's significantly correlated with DNA methylation in 13 types of cancer, but it's correlated with RNA methylation related genes in most cancer. HS6ST2 expression was correlated with immune cell infiltration, immune-related genes, tumor immune microenvironment, and drug resistance in various cancers. Eventually, HS6ST2 was validated in human lung adenocarcinoma tissues. Our study reveals that HS6ST2 can function as a prognostic marker in various malignant tumors because of its role in tumorigenesis and tumor immunity.

Crystal structure of Arabidopsis thaliana sulfotransferase SOT16 involved in glucosinolate biosynthesis

Glucosinolates (GSLs), a class of secondary metabolites found in Brassicaceae plants, play important roles in plant defense and contribute distinct flavors and aromas when used as food ingredients. Following tissue damage, GSLs undergo enzymatic hydrolysis to release bioactive volatile compounds. Understanding GSL biosynthesis and enzyme involvement is crucial for improving crop quality and advancing agriculture. Plant sulfotransferases (SOTs) play a key role in the final step of GSL biosynthesis by transferring sulfate groups to the precursor molecules. In the present study, we investigated the enzymatic reaction mechanism and broad substrate specificity of Arabidopsis thaliana sulfotransferase AtSOT16, which is involved in GSL biosynthesis, using crystal structure analysis. Our analysis revealed the specific catalytic residues involved in the sulfate transfer reaction and supported the hypothesis of a concerted acid-base catalytic mechanism. Furthermore, the docking models showed a strong correlation between the substrates with high predicted binding affinities and those experimentally reported to exhibit high activity. These findings provide valuable insights into the enzymatic reaction mechanisms and substrate specificity of GSL biosynthesis. The information obtained in this study may contribute to the development of novel strategies for manipulating GSL synthesis pathways in Brassica plants and has potential agricultural applications.

Electrodialysis Separation and Selective Concentration of Sulfuric Acid and Nickel Sulfate Mixed Solution Using Membranes Modified by Polyaniline

The paper discusses the electrodialysis treatment of mixed nickel sulfate and sulfuric acid solution using polyaniline surface-modified cation exchange membranes. The modified membranes are obtained on the basis of industrial cation-exchange MK-40 heterogeneous and MF-4SK homogeneous membranes by in situ oxidative polymerization of aniline under electrodialysis conditions. The conductive and diffusion characteristics of the initial and modified membranes in solutions of sulfuric acid and nickel sulfate are studied. It is shown that the modification of membranes with polyaniline leads to a decrease in their electrical conductivity and diffusion permeability while maintaining high selectivity. The diffusion permeability of cation-exchange membranes is higher in nickel sulfate solutions compared to sulfuric acid one, while an inverse dependence is found for anion-exchange membranes. The competitive transfer of sulfuric acid and nickel sulfate during electrodialysis separation and concentration of their mixture using initial and modified cation-exchange membranes paired with anion-exchange MA-41 membrane are studied. It is shown that the applying a polyaniline layer with positively charged groups on the surface of the MK-40 or MF-4SK cation-exchange membranes suppresses the transport of doubly charged nickel ions both in the separation and concentration modes over the entire range of current densities. The greatest repulsion effect is observed for homogeneous modified membranes, where the selective permeability coefficient P(H2SO4/NiSO4) increases from 0.7–1.7 to 32.5–19.7 depending on the current density. It is established that the use of polyaniline modified cation-exchange membranes permits to concentrate the solution containing 0.1 mol-eq/L (4.9 g/L) H2SO4 and 0.1 mol-eq/L (7.7 g/L) NiSO4, with simultaneous separation into sulfuric acid with concentration about 2.4 mol-eq/L (120 g/L) and nickel sulfate solutions. The content of nickel sulfate in the concentrate does not exceed 0.13 mol-eq/L (10 g/L).

Sulfatide-selectin signaling in the spinal cord induces mechanical allodynia.

Sulfatide is a sulfated glycosphingolipid that is present abundantly in myelin sheaths of the brain and spinal cord. It is synthesized by a cerebroside sulfotransferase encoded by Gal3st1, which catalyzes the transfer of sulfate from 3'-phosphoadenylylsulfate to galactosylceramide. We previously reported that Gal3st1 gene expression in the spinal cord is up-regulated 1day after intraplantar injection of complete Freund's adjuvant (CFA), indicating that sulfatide is involved in inflammatory pain. In the present study, we found that intrathecal injection of sulfatide led to mechanical allodynia. Sulfatide caused levels of glial fibrillary acidic protein (GFAP) and nitric oxide in the spinal cord to increase. Mechanical allodynia induced by intrathecal injection of sulfatide was blocked by nitric oxide synthase inhibitors and by suppression of astrocyte activation by L-α-aminoadipate. These results suggest that sulfatide-induced mechanical allodynia involved glial activation and nitric oxide production. Blocking selectin, a sulfatide-binding protein, with bimosiamose attenuated sulfatide-induced allodynia and ameliorated CFA-induced mechanical allodynia during inflammatory pain. Finally, elevated levels of sulfatide concentration in the spinal cord were observed during CFA-induced inflammatory pain. The elevated sulfatide levels enhanced selectin activation in the spinal cord, resulting in mechanical allodynia. Our data suggest that sulfatide-selectin interaction plays a key role in inflammatory pain.

Production of lithium hydroxide by electrodialysis with bipolar membranes

• LiOH (2.2 M) and H 2 SO 4 (1.3 M) were successfully produced from Li 2 SO 4 (1.5 M). • The specific energy consumption was estimated to be around 7 kWh/kg LiOH produced. • The LiOH purity was improved to 99.75%, by limiting the H 2 SO 4 concentration in the acid compartment. • Battery grade LiOH may be possible after only a small reduction of the sulfate content. In conventional lithium extraction processes, highly purified lithium sulfate is generated as an intermediate, which is transformed into lithium hydroxide for lithium-ion battery applications. This study illustrates the technical feasibility of converting the pre-purified lithium sulfate (Li 2 SO 4 ) into lithium hydroxide (LiOH) and sulfuric acid (H 2 SO 4 ) using an electrodialysis with bipolar membrane (EDBM) process. A feed solution of 1.5 mol/L Li 2 SO 4 is converted into 2.2 mol/L LiOH and 1.3 mol/L H 2 SO 4 with a specific energy consumption of less than 10 kWh/kg LiOH produced. Due to the increased acid and base volume caused by electroosmosis, both acid and base concentrations are lower than that estimated based on stoichiometry. Increasing the initial LiOH solution concentration decreases the sulfate transfer from the acid and feed solutions to the base solution. Maintaining a low concentration of H 2 SO 4 in the acid compartment (0.05 mol/L) significantly reduces the migration flux of sulfate ions through the bipolar membrane, leading to a LiOH purity of ~99.75%. This also lowers the energy consumption to ~7 kWh/kg, with a reduced average sulfate leakage flux into the base solution (~0.008 mol/m 2 hr). Battery grade LiOH could be obtained after a small further reduction of the sulfate content. This work demonstrates that EDBM can potentially replace the current chemical precipitation technique to produce LiOH from Li 2 SO 4 , by eliminating the use of a large quantity of alkali and the associated precipitation and solid–liquid separation steps.

The Competitive Adsorption of Chromate and Sulfate on Ni-Substituted Magnetite Surfaces: An ATR-FTIR Study

With similar chemical properties and geometrical configurations, sulfate and chromate display interesting competitive adsorption on mineral surfaces. Although such issues have been investigated on several Fe (hydr)oxide surfaces, e.g., ferrihydrite, goethite and hematite, the competitive adsorption on magnetite surfaces and the constraint mechanism have seldom been studied. This impedes the understanding of the transfer and fate of chromate and sulfate on magnetite surfaces, as magnetite is not only a useful adsorbent but also an efficient reductant to decrease the mobility and toxicity of chromium. In the present study, the geometries of the competitive adsorption of chromate and sulfate on Ni-substituted magnetite surfaces over a pH range of 4–9 were investigated using in situ attenuated total reflectance Fourier transform infrared spectroscopy and two-dimensional correlation analysis. In individual adsorption, nonprotonated monodentate mononuclear (NMM) complexes dominated chromate adsorption, accompanied by a few bidentate binuclear (BB) complexes. For sulfate, NMM complexes and outer-sphere (OS) species predominated under acidic and neutral–alkaline conditions, respectively. The above variation in adsorption configuration resulted in the different adsorption competitiveness between chromate and sulfate at different pH values. Specifically, the NMM complexes of chromate were substituted by NMM sulfate complexes under acidic conditions and vice versa. However, under neutral and alkaline conditions, the OS species of sulfate scarcely affected the adsorption of chromate. The adsorption affinity of chromate and sulfate on Ni-substituted magnetite increased in the following order: OS complex (sulfate) < NMM complexes (chromate) < NMM complexes (sulfate).

Redox transfer at subduction zones: insights from Fe isotopes in the Mariana forearc

Subduction zones are active sites of chemical exchange between the Earth’s surface and deep interior and play a fundamental role in regulating planet habitability. However, the mechanisms by which redox sensitive elements (e.g., iron, carbon and sulfur) are cycled during subduction remains unclear. Here we use Fe stable isotopes (δ56Fe), which are sensitive to redox-related processes, to examine forearc serpentinite clasts recovered from deep sea drilling of mud volcanoes formed above the Mariana subduction zone in the Western Pacific. We show that serpentinisation of the forearc by slab-derived fluids produces dramatic δ56Fe variation. Unexpected negative correlations between serpentinite bulk δ56Fe, fluid-mobile element concentrations (e.g., B, As) and Fe3+/ƩFe suggest a concomitant oxidation of the mantle wedge through the transfer of isotopically light iron by slab-derived fluids. This process must reflect the transfer of either sulfate- or carbonate-bearing fluids that preferentially complex isotopically light Fe.

Directed aryl sulfotransferase evolution toward improved sulfation stoichiometry on the example of catechols.

Sulfation is an important way for detoxifying xenobiotics and endobiotics including catechols. Enzymatic sulfation occurs usually with high chemo- and/or regioselectivity under mild reaction conditions. In this study, a two-step p-NPS-4-AAP screening system for laboratory evolution of aryl sulfotransferase B (ASTB) was developed in 96-well microtiter plates to improve the sulfate transfer efficiency toward catechols. Increased transfer efficiency and improved sulfation stoichiometry are achieved through the two-step screening procedure in a one-pot reaction. In the first step, the p-NPS assay is used (detection of the colorimetric by-product, p-nitrophenol) to determine the apparent ASTB activity. The sulfated product, 3-chlorocatechol-1-monosulfate, is quantified by the 4-aminoantipyrine (4-AAP) assay in the second step. Comparison of product formation to p-NPS consumption ensures successful directed evolution campaigns of ASTB. Optimization yielded a coefficient of variation below 15% for the two-step screening system (p-NPS-4-AAP). In total, 1760 clones from an ASTB-SeSaM library were screened toward the improved sulfation activity of 3-chlorocatechol. The turnover number (kcat = 41 ± 2 s-1) and catalytic efficiency (kcat/KM = 0.41 μM-1 s-1) of the final variant ASTB-M5 were improved 2.4- and 2.3-fold compared with ASTB-WT. HPLC analysis confirmed the improved sulfate stoichiometry of ASTB-M5 with a conversion of 58% (ASTB-WT 29%; two-fold improvement). Mass spectrometry (MS) and nuclear magnetic resonance spectroscopy (NMR) confirmed the chemo- and regioselectivity, which yielded exclusively 3-chlorocatechol-1-monosulfate. For all five additionally investigated catechols, the variant ASTB-M5 achieved an improved kcat value of up to 4.5-fold and sulfate transfer efficiency was also increased (up to 2.3-fold).

Transition-State Interactions in a Promiscuous Enzyme: Sulfate and Phosphate Monoester Hydrolysis by Pseudomonas aeruginosa Arylsulfatase.

Pseudomonas aeruginosa arylsulfatase (PAS) hydrolyzes sulfate and, promiscuously, phosphate monoesters. Enzyme-catalyzed sulfate transfer is crucial to a wide variety of biological processes, but detailed studies of the mechanistic contributions to its catalysis are lacking. We present linear free energy relationships (LFERs) and kinetic isotope effects (KIEs) of PAS and analyses of active site mutants that suggest a key role for leaving group (LG) stabilization. In LFERs PASWT has a much less negative Brønsted coefficient (βleavinggroupobs-Enz = -0.33) than the uncatalyzed reaction (βleavinggroupobs = -1.81). This situation is diminished when cationic active site groups are exchanged for alanine. The considerable degree of bond breaking during the transition state (TS) is evidenced by an 18Obridge KIE of 1.0088. LFER and KIE data for several active site mutants point to leaving group stabilization by active site K375, in cooperation with H211. 15N KIEs and the increased sensitivity to leaving group ability of the sulfatase activity in neat D2O (ΔβleavinggroupH-D = +0.06) suggest that the mechanism for S-Obridge bond fission shifts, with decreasing leaving group ability, from charge compensation via Lewis acid interactions toward direct proton donation. 18Ononbridge KIEs indicate that the TS for PAS-catalyzed sulfate monoester hydrolysis has a significantly more associative character compared to the uncatalyzed reaction, while PAS-catalyzed phosphate monoester hydrolysis does not show this shift. This difference in enzyme-catalyzed TSs appears to be the major factor favoring specificity toward sulfate over phosphate esters by this promiscuous hydrolase, since other features are either too similar (uncatalyzed TS) or inherently favor phosphate (charge).

Downregulation of HS6ST2 by miR-23b-3p enhances matrix degradation through p38 MAPK pathway in osteoarthritis

Osteoarthritis (OA) is the most common form of arthritis involving major structural changes of peripheral joints and local or systemic inflammation and in lack of therapeutic approaches because of complexity of underlying molecular basis. Our previous work showed that HS6ST2, an enzyme involved in the transfer of sulfate, is downregulated in cartilage tissues of OA patients compared with normal donors, but little is known about its regulatory mechanism. In this study, we demonstrated that the expression of HS6ST2 was lower in OA-damaged cartilage than smooth cartilage from the same patient. In chondrocytes, HS6ST2 could be targeted by miR-23b-3p, which was higher expressed in OA-damaged cartilage. Under TNF-α stimulation, the expression of HS6ST2 was found inversely correlated with the expression of miR-23b-3p. Downregulation of HS6ST2 regulated by overexpression of miR-23b-3p and siRNAs against HS6ST2 could enhance the protein level of MMP13 and aggravate the matrix degradation in chondrocytes. Increased expression of MMP13 depended on activity of p38 MAPK rather than total p38 MAPK level and was abrogated by HS6ST2 overexpression. Together, the results indicated that downregulated HS6ST2 targeted by miR-23b-3p promotes matrix degradation by activating p38 MAPK in chondrocytes and OA cartilage.

Carbonate Transfer during the Onset of Slab Devolatilization: New Insights from Fe and Zn Stable Isotopes

Long-term carbon cycling is a subject of recent controversy as new mass balance calculations suggest that most carbon is transferred from the slab to the mantle wedge by fluids during subduction, limiting the efficiency of carbon recycling to the deep mantle. Here, we examine the large scale mobility of carbon during subduction using new isotopic tracers sensitive to H–C–O–S–Cl fluids, namely iron and zinc stable isotopes, in samples interpreted to represent residual slab (Queyras, Western Alps) and sub-arc mantle (Kohistan, Himalaya). We show that during subduction there are several stages of carbonate precipitation and dissolution at metasomatic interfaces between metasedimentary and ultramafic rocks in the slab. During the early stages of subduction, before the slab reaches the 300–400C isotherms, the infiltration of sediment-derived fluids into ultramafic lithologies enhances carbonate precipitation in antigorite-bearing serpentinites. Carbonate storage in serpentinites, therefore, acts as a temporary reservoir of carbon in subduction zones. This episode is accompanied by a decrease in serpentinite iron isotope composition (d56Fe), due to interaction with low-d56Fe sediment-derived fluids, and an increase in the concentrations of fluid-mobile elements (e.g. B, Li, As). At higher temperatures (>400C), carbonate is leached from the serpentinites by fluids. This is accompanied by a decrease in serpentinite zinc isotope composition (d66Zn) which we interpret as the release of a carbonate-bearing fluid with an isotopically heavy d66Zn signature. Thermodynamic modelling shows that the sudden change in fluid carbon mobility is due to a decrease in the aCO2 of the fluids released during slab prograde metamorphism, which shifts from sediment- to serpentinite-dominated dehydration. This demonstrates that slab fluids bearing oxidized carbon (e.g. CO2), associated with isotopically light Fe, heavy Zn and fluid-mobile elements, can be released before the slab reaches eclogite facies P-T conditions. These observations provide strong evidence for the mobility of carbon in fluids during the early stages of subduction. Moreover, the fluids released will act as a potential metasomatic agent for the fore-arc mantle (or slab/mantle interface). The observation of carbonate-bearing metamorphic veins in the Himalayan sub-arc mantle with complementary light d56Fe and heavy d66Zn signatures provides further support for the large scale transfer of both sulphate- and carbonate-bearing fluids during the early stages of subduction. This suggests that the fore-arc may have an important role in delivering water, sulfur and carbon to the source of arc-magmas.

Homozygous CHST11 mutation in chondrodysplasia, brachydactyly, overriding digits, clino-symphalangism and synpolydactyly

BackgroundCarbohydrate sulfotransferase 11 (CHST11) is a membrane protein of Golgi that catalyses the transfer of sulfate to position 4 of the N-acetylgalactosamine residues of chondroitin. Chondroitin sulfate is the predominant...

Evaluation of interfacial sulfate complexation by a bis-thiourea ionophore at water-organic interfaces using microelectrochemistry and high resolution mass spectrometry

Simple, fast and low cost methods for the detection of sulfate are required for different applications. Electrochemistry at water/o-nitrophenyloctylether (W/NPOE) interfaces was employed to evaluate sulfate detection by ionophore-facilitated ion-transfer at an array of micro-interfaces. With ionophore 1,3-[Bis(3-phenylthioureidomethyl)]benzene present in the NPOE phase, the transfer of sulfate across the interface was determined by voltammetry at ca. −0.35V for 0.01M Na2SO4 on the Galvani potential scale. The potentiometric detection limit for sulfate was 0.6×10−6M, based on the shift in the half-wave transfer potential with concentration. Amperometric detection limits for forward and reverse ion transfer currents were determined to be 14×10−6M and 0.8×10−6M, respectively. Electrochemical analysis of the half-wave potential versus logCSO42−w and the corresponding electrospray ionisation – high resolution mass spectrometry (ESI-HRMS) analysis of W/NPOE emulsions indicated interfacial complexation via the formation of 1:1 sulfate:ionophore complexes. ESI-HRMS analysis of W/NPOE emulsions formed with water samples from an advanced water treatment plant revealed the binding of the ionophore to potential interferences from this environment, thus providing a guide to sensor development.

Promiscuity in the Enzymatic Catalysis of Phosphate and Sulfate Transfer.

The enzymes that facilitate phosphate and sulfate hydrolysis are among the most proficient natural catalysts known to date. Interestingly, a large number of these enzymes are promiscuous catalysts that exhibit both phosphatase and sulfatase activities in the same active site and, on top of that, have also been demonstrated to efficiently catalyze the hydrolysis of other additional substrates with varying degrees of efficiency. Understanding the factors that underlie such multifunctionality is crucial both for understanding functional evolution in enzyme superfamilies and for the development of artificial enzymes. In this Current Topic, we have primarily focused on the structural and mechanistic basis for catalytic promiscuity among enzymes that facilitate both phosphoryl and sulfuryl transfer in the same active site, while comparing this to how catalytic promiscuity manifests in other promiscuous phosphatases. We have also drawn on the large number of experimental and computational studies of selected model systems in the literature to explore the different features driving the catalytic promiscuity of such enzymes. Finally, on the basis of this comparative analysis, we probe the plausible origins and determinants of catalytic promiscuity in enzymes that catalyze phosphoryl and sulfuryl transfer.

CHST11/13 Regulate the Metastasis and Chemosensitivity of Human Hepatocellular Carcinoma Cells Via Mitogen-Activated Protein Kinase Pathway.

Carbohydrate sulfotransferases 11-13 (CHST11-13), that catalyze the transfer of sulfate to position 4 of the GalNAc residue of chondroitin, have been implicated in various diseases. This study aimed to clarify the association of CHST11-13 expression with metastasis and drug sensitivity in hepatocellular carcinoma (HCC) cells. We measured the levels of CHST11 and CHST13 in a series of HCC cells using real-time PCR and Western blotting. After RNAi and forced expression treatment of CHST11 and CHST13 in MHCC97L and MHCC97H cells, metastatic potential and drug sensitivity of the two cells were investigated with ECM invasion assay, drug sensitivity assay, and in vivo antitumor activity assay. By real-time PCR and Western blotting, we explored the possible impacts of these two genes on mitogen-activated protein kinase (MAPK) signal pathway. MAPK pathway was blocked by PD98059 or SP600125 to elucidate the effects of MAPK pathway on metastasis and chemosensitivity. Significantly reduced levels of CHST11 and CHST13 were observed in highly invasive MHCC97H cells compared with those of MHCC97L cell line with low metastatic potential. Decreased or forced expression of CHST11 and CHST13 altered metastatic potential and drug sensitivity of MHCC97L and MHCC97H cells. Remarkable alteration of MAPK activity was shown in two HCC cells with genetic manipulation. Conversely, pharmacologic inhibition of the MAPK pathway suppressed invasive potential and rescued drug sensitivity of MHCC97H cells. Our results have demonstrated that CHST11 and CHST13 negatively modulate metastasis and drug resistance of HCC cells probably via oncogenic MAPK signal pathway.

Isotopic evidence for iron mobility during subduction

Subduction zones are one of the most important sites of chemical interchange between the Earth’s surface and interior. One means of explaining the high Fe3+/ΣFe ratios and oxidized nature of primary arc magmas is the transfer of sulfate (SOX), carbonate (CO3–), and/or iron (Fe3+) bearing fluids from the slab to the overlying mantle. Iron mobility and Fe stable isotope fractionation in fluids are influenced by Fe redox state and the presence of chlorine and/or sulfur anions. Here we use Fe stable isotopes (δ56Fe) as a tracer of iron mobility in serpentinites from Western Alps metaophiolites, which represent remnants of oceanic lithosphere that have undergone subduction-related metamorphism and devolatilization. A negative correlation (R2 = 0.72) is observed between serpentinite bulk δ56Fe and Fe3+/ΣFe that provides the first direct evidence for the release of Fe-bearing fluids during serpentinite devolatilization in subduction zones. The progressive loss of isotopically light Fe from the slab with increasing degree of prograde metamorphism is consistent with the release of sulfate-rich and/or hypersaline fluids, which preferentially complex isotopically light Fe in the form of Fe(II)-SOX or Fe(II)-Cl2 species. Fe isotopes can therefore be used as a tracer of the nature of slab-derived fluids.

Pharmacokinetics and placental transfer of magnesium sulfate in pregnant women

Magnesium sulfate is one of the most commonly prescribed intravenous medications in obstetrics. Despite its widespread use, there are limited data about magnesium pharmacokinetics, and magnesium is prescribed empirically without dose adjustment for different indications. The aim of this study was to characterize the pharmacokinetics and placental transfer of magnesium sulfate in pregnant women and to determine key covariates that impact the pharmacokinetics. This is a prospective pharmacokinetic cohort study ofpregnant women who were prescribed magnesium sulfate for preeclampsia, preterm labor, or extreme prematurity. Women received a 4-g loading dose and 2 g/h maintenance dose as clinically indicated. Maternal blood samples were obtained before and at multiple time points during and after magnesium administration. Cord blood also was sampled at delivery. A population pharmacokinetic approach that used a nonlinear mixed-effects modeling was used to characterize magnesium disposition. Pharmacokinetic profiles of 111 pregnant women were analyzed. Magnesium clearance was 3.98 L/h in preeclamptic women and5.88 L/h non-preeclamptic women. Steady-state concentration ofmagnesium was 7.2 mg/dL in preeclamptic women compared with 5.1mg/dL in non-preeclamptic women. Maternal weight significantly impacted time to steady state. The ratio of the mean umbilical vein magnesium level to the mean maternal serum magnesium level at the time of delivery was 0.94 ± 0.15. The study accurately characterizes the pharmacokinetics of magnesium administered to pregnant women. Preeclamptic status and maternal weight significantly impact serum magnesium levels. This pharmacokinetic model could be applied to larger cohorts to help tailor magnesium treatment and account for these covariates.