Only in Titles

Search results for: Recombinant Human GSTO1 Proteins


#24111988   2013/10/28 To Up

A competitive chemical-proteomic platform to identify zinc-binding cysteines.

Zinc ions (Zn(2+)) play vital catalytic, structural, and regulatory roles in protein function and are commonly chelated to cysteine residues within the protein framework. Current methods to identify Zn(2+)-binding cysteines rely on computational studies based on known Zn(2+)-chelating motifs, as well as high-resolution structural data. These available approaches preclude the global identification of putative Zn(2+)-chelating cysteines, particularly on poorly characterized proteins in the proteome. Herein, we describe an experimental platform that identifies metal-binding cysteines on the basis of their reduced nucleophilicity upon treatment with metal ions. As validation of our platform, we utilize a peptide-based cysteine-reactive probe to show that the known Zn(2+)-chelating cysteine in sorbitol dehydrogenase (SORD) demonstrates an expected loss in nucleophilicity in the presence of Zn(2+) ions and a gain in nucleophilicity upon treatment with a Zn(2+) chelator. We also identified the active-site cysteine in glutathione S-transferase omega-1 (GSTO1) as a potential Zn(2+)-chelation site, albeit with lower metal affinity relative to SORD. Treatment of recombinant GSTO1 with Zn(2+) ions results in a dose-dependent decrease in GSTO1 activity. Furthermore, we apply a promiscuous cysteine-reactive probe to globally identify putative Zn(2+)-binding cysteines across ∼900 cysteines in the human proteome. This proteomic study identified several well-characterized Zn(2+)-binding proteins, as well as numerous uncharacterized proteins from functionally distinct classes. This platform is highly versatile and provides an experimental tool that complements existing computational and structural methods to identify metal-binding cysteine residues.
Nicholas J Pace, Eranthie Weerapana

2860 related Products with: A competitive chemical-proteomic platform to identify zinc-binding cysteines.

100ug Lyophilized 1 G100 assays200 ug 100ul0.1 mg1 mg1 module0.2 mg1 module0.1 mg100 tests

Related Pathways


#23827461   2013/07/01 To Up

Systematic identification and characterization of glutathione S-transferases in cynomolgus macaque.

Glutathione S-transferases (GSTs) are essential drug-metabolizing enzymes, involved in conjugation of various endogenous and exogenous substrates. Cynomolgus macaque is an important primate species in drug metabolism studies; however, cynomolgus GSTs have not been fully characterized. In this study the cDNAs of 12 GSTs (GSTA3-A5, GSTK1, GSTM2-M4, GSTO2, GSTP1, GSTS1, and GSTT1/2) were isolated from cynomolgus macaque and rhesus macaque liver. Cynomolgus GSTM1 cDNA was not amplified and only an aberrantly spliced GSTM1 transcript was isolated from rhesus macaque. Amino acid sequences of these 12 GSTs shared high sequence identities (93-98%) and were clustered into the same clades as the human orthologs in the phylogenetic tree. The 12 GSTs had exon-intron structures similar to the human orthologs, and exhibited distinct tissue expression patterns. GSTA3, GSTA5, and GSTM3/O2 were expressed predominantly in adrenal gland, jejunum, and testis, respectively, whereas the other GSTs showed universal expression patterns in the 10 tissues analyzed. Comparison of expression levels showed that GSTA1, GSTK1, GSTA3, and GSTM3 were most abundantly expressed in liver/jejunum, kidney, adrenal gland, and testis, respectively. Metabolic assays of proteins expressed heterologously in Escherichia coli, showed that all 12 GSTs and 5 previously identified GSTs, GSTA1/2, GSTM5, GSTO1, and GSTZ1, catalyzed the conjugation of GST substrate(s) 1-chloro-2,4-dinitrobenzene and/or 1,2-epoxy-3-(p-nitrophenoxy)propane, indicating that these 17 GSTs are functional drug-metabolizing enzymes. These results suggest that the 12 GST genes examined in this study are expressed and encoded functional enzymes in cynomolgus macaque.
Yasuhiro Uno, Norie Murayama, Mutsuki Kunori, Hiroshi Yamazaki

2430 related Products with: Systematic identification and characterization of glutathione S-transferases in cynomolgus macaque.

100 μg100 μg100 μg1 Set8 Sample Kit100 μg1 Set100100 μg2ug

Related Pathways


#21899313   2011/09/27 To Up

Potent and selective inhibitors of glutathione S-transferase omega 1 that impair cancer drug resistance.

Glutathione S-transferases (GSTs) are a superfamily of enzymes that conjugate glutathione to a wide variety of both exogenous and endogenous compounds for biotransformation and/or removal. Glutathione S-tranferase omega 1 (GSTO1) is highly expressed in human cancer cells, where it has been suggested to play a role in detoxification of chemotherapeutic agents. Selective inhibitors of GSTO1 are, however, required to test the role that this enzyme plays in cancer and other (patho)physiological processes. With this goal in mind, we performed a fluorescence polarization activity-based protein profiling (fluopol-ABPP) high-throughput screen (HTS) with GSTO1 and the Molecular Libraries Small Molecule Repository (MLSMR) 300K+ compound library. This screen identified a class of selective and irreversible α-chloroacetamide inhibitors of GSTO1, which were optimized to generate an agent KT53 that inactivates GSTO1 with excellent in vitro (IC(50) = 21 nM) and in situ (IC(50) = 35 nM) potency. Cancer cells treated with KT53 show heightened sensitivity to the cytotoxic effects of cisplatin, supporting a role for GSTO1 in chemotherapy resistance.
Katsunori Tsuboi, Daniel A Bachovchin, Anna E Speers, Timothy P Spicer, Virneliz Fernandez-Vega, Peter Hodder, Hugh Rosen, Benjamin F Cravatt

2901 related Products with: Potent and selective inhibitors of glutathione S-transferase omega 1 that impair cancer drug resistance.

100ul 100ul100ul100ul100ul1x96 well plate100ul100ul100ul100ug100ug Lyophilized10 mg

Related Pathways


#21311795   2011/02/10 To Up

Fluorescent labeling of membrane proteins on the surface of living cells by a self-catalytic glutathione S-transferase omega 1 tag.

Imaging a specific protein of interest in live cell has versatile applications in biological research. Recently, diverse chemical tags have been developed to overcome the limits of autofluorescence protein (FP) tags. However, current chemical methods still need to be progressed to compete with FPs in the scope of specificity and convenience in staining procedure. We report a novel protein tagging method that provides a convenient and specific fluorescent labeling of membrane proteins. Ω tag is developed by employing a mammalian enzyme glutathione sulfur-transferase omega 1 (GSTO1) and its partner dye, 5-bromomethyl fluorescein (BMF). Ω-tagged membrane proteins were labeled by BMF efficiently for live cell imaging and in-gel analysis. Endocytosis of epidermal growth factor receptor (EGFR) was successfully visualized by using this Ω tagging system. Ω tag will provide a convenient tool for the physiological study of membrane proteins in live cells.
Jae-Jung Lee, Jyunghyun Son, Hyung-Ho Ha, Young-Tae Chang

1579 related Products with: Fluorescent labeling of membrane proteins on the surface of living cells by a self-catalytic glutathione S-transferase omega 1 tag.

10reactions 10reactions 10reactions 10reactions 10reactions (20 µl each)10reactions

Related Pathways


#21106529   2010/11/24 To Up

Novel folding and stability defects cause a deficiency of human glutathione transferase omega 1.

The polymorphic deletion of Glu-155 from human glutathione transferase omega1 (GSTO1-1) occurs in most populations. Although the recombinant ΔGlu-155 enzyme expressed in Escherichia coli is active, the deletion causes a deficiency of the active enzyme in vivo. The crystal structure and the folding/unfolding kinetics of the ΔGlu-155 variant were determined in order to investigate the cause of the rapid loss of the enzyme in human cells. The crystal structure revealed altered packing around the Glu-155 deletion, an increase in the predicted solvent-accessible area and a corresponding reduction in the buried surface area. This increase in solvent accessibility was consistent with an elevated Stern-Volmer constant. The unfolding of both the wild type and ΔGlu-155 enzyme in urea is best described by a three-state model, and there is evidence for the more pronounced population of an intermediate state by the ΔGlu-155 enzymes. Studies using intrinsic fluorescence revealed a free energy change around 14.4 kcal/mol for the wild type compared with around 8.6 kcal/mol for the ΔGlu-155 variant, which indicates a decrease in stability associated with the Glu-155 deletion. Urea induced unfolding of the wild type GSTO1-1 was reversible through an initial fast phase followed by a second slow phase. In contrast, the ΔGlu-155 variant lacks the slow phase, indicating a refolding defect. It is possible that in some conditions in vivo, the increased solvent-accessible area and the low stability of the ΔGlu-155 variant may promote its unfolding, whereas the refolding defect limits its refolding, resulting in GSTO1-1 deficiency.
Huina Zhou, Joseph Brock, Marco G Casarotto, Aaron J Oakley, Philip G Board

1266 related Products with: Novel folding and stability defects cause a deficiency of human glutathione transferase omega 1.

100ul 100ul100ul 100ul200 0.1 mg100 μg1000 100ul1 ml0.1ml (1mg/ml)

Related Pathways


#17019444   2006/10/01 To Up

Tocopherol esters inhibit human glutathione S-transferase omega.

Human glutathione S-transferase omega 1-1 (hGSTO1-1) is a newly identified member of the glutathione S-transferase (GST) family of genes, which also contains alpha, mu, pi, sigma, theta, and zeta members. hGSTO1-1 catalyzes the reduction of arsenate, monomethylarsenate (MMA(V)), and dimethylarsenate (DMA(V)) and exhibits thioltransferase and dehydroascorbate reductase activities. Recent evidence has show that cytokine release inhibitory drugs, which specifically inhibit interleukin-1b (IL-1b), directly target hGSTO1-1. We found that (+)-alpha-tocopherol phosphate and (+)-alpha-tocopherol succinate inhibit hGSTO1-1 in a concentration-dependent manner with IC50 values of 2 microM and 4 microM, respectively. A Lineweaver-Burk plot demonstrated the uncompetitive nature of this inhibition. The molecular mechanism behind the inhibition of hGSTO1-1 by alpha-tocopherol esters (vitamin E) is important for understanding neurodegenerative diseases, which are also influenced by vitamin E.
Adriana Sampayo-Reyes, Robert A Zakharyan

1697 related Products with: Tocopherol esters inhibit human glutathione S-transferase omega.

100ul 100ul0.5mg10 100ul96 wells (1 kit)96T100μg1mg5 0.1 ml

Related Pathways


#16781109   2006/05/24 To Up

Inhibition of human glutathione S-transferase omega by tocopherol succinate.

Glutathione S-transferases (GSTs) are a family of multifunctional enzymes that are present in all living organisms. Their main function is the detoxification of electrophilic compounds. Glutathione conjugation is the major detoxification pathway available to the organism to trap toxic substances. Based on their substrate specificity, sequence structure, catalytic activity, immunogenicity and sensitivity to inhibitors, the mammalian GSTs form seven distinct classes termed alpha, mu, pi, sigma, theta, zeta, and new class of human GSTs designated omega. Human GST omega 1-1 (hGSTO1-1) is identical to human monomethylarsenic acid (MMAV), the rate-limiting enzyme for biotransformation of inorganic arsenic. It is expressed in a wide range of human tissues, including brain. Several studies have indicated a role for an Omega-class GST gene in the early onset of both Alzheimer's and Parkinson's diseases, and it is possible that hGSTO1-1 may be involved in the modulation of the activity of interleukin-1 (IL-1) which play a major role in a wide range of inflammatory disease. Compounds that target IL-1 production are being investigated. We found that (+)-alpha-tocopherol succinate inhibited the reduction monomethylarsenate (MMAV) and dimethylarsenate (DMAV) in a concentration-dependent manner with an IC(50) of 4 and 3 microM, respectively. The kinetics indicated an uncompetitive inhibition of the MMA(V) and DMA(V) reducing activity of hGSTO1-1.
A Sampayo-Reyes, R A Zakharyan

2191 related Products with: Inhibition of human glutathione S-transferase omega by tocopherol succinate.

100ul 100ul0.5mg 100ul 100ul1 mg96 wells (1 kit)0.1ml (1mg/ml)100 μg500 100ul

Related Pathways


Error loading info... Pleas try again later.

#16097802   // To Up

Interactions of sodium selenite, glutathione, arsenic species, and omega class human glutathione transferase.

Human monomethylarsenate reductase [MMA(V) reductase] and human glutathione S-transferase omega 1-1 (hGSTO1-1) [because MMA(V) reductase and hGSTO1-1 are identical proteins, the authors will utilize the designation "hGSTO1-1"] are identical proteins that catalyze the reduction of arsenate, monomethylarsenate [MMA(V)], and dimethylarsenate [DMA(V)]. Sodium selenite (selenite) inhibited the reduction of each of these substrates by the enzyme in a concentration-dependent manner. The kinetics indicated a noncompetitive inhibition of the MMA(V), DMA(V), or arsenate reducing activity of hGSTO1-1. The inhibition of the MMA(V) reducting activity of hGSTO1-1 by selenite was reversed by 1 mM DL-dithiothreitol (DTT) but not by reduced glutathione (GSH), which is a required substrate for the enzyme. Neither superoxide anion nor hydrogen peroxide was involved in the selenite inhibition of hGSTO1-1. MALDI-TOF and MS/MS analysis demonstrated that five molecules of GSH were bound to one monomer of hGSTO1-1. Four of the five cysteines of the monomer were glutathionylated. Cys-32 in the active center, however, exists mostly in the sulfhydryl form since it was alkylated consistently by iodoacetamide. MALDI-TOF mass spectra analysis of hGSTO1-1 after reaction with GSH and sodium selenite indicated that selenium was integrated into hGSTO1-1 molecules. Three selenium were found to be covalently bonded to the monomer of hGSTO1-1 with three molecules of GSH. It is proposed that the reaction products of the reduction of selenite inhibited the activity of hGSTO1-1 by reacting with disulfides of glutathionylated cysteines to form bis (S-cysteinyl)selenide and S-selanylcysteine and had little or no interaction with the sulfhydryl of Cys-32 in the active site of the enzyme.
Robert A Zakharyan, George Tsaprailis, Uttam K Chowdhury, Alba Hernandez, H Vasken Aposhian

1635 related Products with: Interactions of sodium selenite, glutathione, arsenic species, and omega class human glutathione transferase.

100ul 100ul 100ul 100ul500 0.5mg0.1ml (1mg/ml) 100ul 100ul100 μg1x96 well plate1 mg

Related Pathways


#15942673   // To Up

Cloning, expression and characterization of human glutathione S-transferase Omega 2.

The class of Omega glutathione transferases is newly identified with novel structural and functional characteristics. Human GSTO 1-1 (glutathione S-transferase Omega 1) is the first member of the GST Omega class. It was found to play a role in apoptosis and be in association with age-at-onset of AD and PD. In order to improve the understanding of the properties of other Omega class members, we screened a human fetal brain cDNA library and obtained the human GSTO2 (glutathione S-transferase Omega 2) cDNA. The full-length cDNA of human GSTO2 is 1179 bp long and encodes a protein of 243 amino acid residues. Expression pattern analysis revealed that GSTO2 was ubiquitously expressed at a low level, with a higher expression in pancreas and prostate. Enzyme assays showed that GSTO2 protein had activities similar to Omega class GSTs. It has detectable glutathione-dependent thiol transferase activity and glutathione-dependent dehydroascorbate reductase activity. But different from GSTO1-1, GSTO2 exhibits a high catalytic activity with CDNB. Subcellular localization analysis of GSTO2-EGFP fusion protein revealed that GSTO2 distributed to cytoplasm of COS-7 cells and both cytoplasm and nucleus of L-02, QGY-7703 and SMMC-7721 cells. Overexpression of GSTO2 induced apoptosis of L-02 cells detected by Annexin V-PE staining. The results suggest that GSTO2 may play an important role in cellular signaling.
Liu Wang, Jian Xu, Chaoneng Ji, Shaohua Gu, You Lv, Shi Li, Yiren Xu, Yi Xie, Yumin Mao

2717 related Products with: Cloning, expression and characterization of human glutathione S-transferase Omega 2.

100ul 100ul96 wells (1 kit)0.5mg 100ul200 100ul1000 2x 100ug 100ul100ul

Related Pathways