Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: UMLS:C0272170 (SDS)
 50,377 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Thermal decomposition by the azo initiator 2,2' azobis(2-amidinopropane) dihydrochloride (AAPH) has been widely used as a water-soluble source of free radical initiators capable of inducing lipid peroxidation and protein damage. Here, in a lipid-free system, AAPH alone (40 mM) rapidly induced protein modification and inactivation of the enzyme catalase (EC 1.11.1.6). Using SDS-PAGE, it was shown that protein band intensity is dramatically reduced after 4 h of incubation with AAPH, leading to protein aggregation. Several antioxidants including melatonin, glutathione (GSH) and trolox prevented catalase modification when used at a 250 microM concentration whereas ascorbate was only effective at 1 mM concentration. All the antioxidants tested reduced carbonyl formation although melatonin was the most effective in this regard. Enzyme inactivation caused by AAPH was also significantly reduced by the antioxidants and again melatonin was more efficient than the other antioxidants used in this study. Results shown here demonstrate that alkyl peroxyl radicals inactivate catalase and reduce the effectiveness of cells to defend against free radical damage; the damage to catalase can be prevented by antioxidants, especially melatonin.
 ...
PMID:Oxidative damage to catalase induced by peroxyl radicals: functional protection by melatonin and other antioxidants. 1279 76

A glutathione S-transferase (GST) was purified 266-fold from adult workers of the red imported fire ant, Solenopsis invicta (Hymenoptera: Formicidae) by affinity chromatography and preparative isoelectric focusing. The purified enzyme appeared as a single band on SDS-PAGE and had a Mr of 25.5 kDa. Steady state kinetics assays of the enzyme with 1-chloro-2,4-dinitrobenzene as substrate were conducted. The Vmax, Km CDNB, Km GSH, kcat, kcat/Km CDNB, and kcat/Km GSH for the purified fire ant GST were 87.4 micromol/min/mg, 0.13 mM, 0.84 mM, 74.5 s(-1), 573.1 mM(-1) s(-1), and 88.7 mM(-1) s(-1), respectively. An internal fragment of the enzyme, released by endoproteinase Lys-C digestion, was sequenced and used to design a degenerate oligonucleotide primer. Purified cDNA from a lambda-phage expression library produced from worker fire ants was used as template to amplify a fragment of the GST transcript which was subsequently cloned and sequenced. 5' and 3' rapid amplification of cDNA ends were subsequently conducted after cDNA production by RT-PCR of mRNA from adult worker fire ants. An open reading frame comprising 202 amino acids with a calculated molecular weight of 23,433 and a theoretical pI of 7.84 was located within the transcript beginning at nucleotide 85 and terminating in a stop codon at nucleotide 691. The transcript contained 5' and 3' untranslated regions of 84 and 296 nucleotides, respectively. The sequences of the internal fragments from the purified fire ant GST were identical to corresponding translated regions of the transcript (nucleotides 214-258, and 457-477). Comparison of the deduced amino acid sequence with GSTs from other species revealed that the enzyme was most closely related to sigma class GSTs.
 ...
PMID:Purification, biochemical characterization, and cDNA cloning of a glutathione S-transferase from the red imported fire ant, Solenopsis invicta. 1450 91

Oxidative stress has been considered to be a major cause of cellular injuries in a variety of chronic health problems, such as carcinogenesis and neurodegenerative disorders. The brain appears to be more susceptible to oxidative damage than other organs. Therefore, the existence of antioxidants may be essential in brain protective systems. The antioxidative and free radical scavenging effects of endomorphin 1 (EM1) and endomorphin 2 (EM2), endogenous opioid peptides in the brain, have been investigated in vitro. The oxidative damage was initiated by a water-soluble initiator 2,2'-azobis(2-amidinopropane hydrocholoride) (AAPH) and hydrogen peroxide (H2O2). The linoleic acid peroxidation, DNA and protein damage were monitored by formation of hydroperoxides, by plasmid pBR 322 DNA nicking assay and single-cell alkaline electrophoresis, and by SDS-polyacrylamide gel electrophoresis. Endomorphins can inhibit lipid peroxidation, DNA strand breakage, and protein fragmentation induced by free radical. Endomorphins also reacted with galvinoxyl radicals in homogeneous solution, and the pseudo-first-order rate constants were determined spectrophotometrically by following the disappearance of galvinoxyl radicals. In all assay systems, EM1 was more potent than EM2 and GSH, a major intracellular water-soluble antioxidant. We propose that endomorphins are one of the protective systems against free radical-induced damage in the brain.
 ...
PMID:Endomorphins, endogenous opioid peptides, provide antioxidant defense in the brain against free radical-induced damage. 1463 51

The glutathionylation of human lens proteins was examined by Western-blot analysis with an anti-GSH antibody and scanning. Several different glutathionylated proteins were observed, and a 47 kDa band was of particular interest. This band did not appear after SDS/PAGE under reducing conditions, suggesting that it was a glutathionylated fraction. The 47 kDa band was found principally in the outer part of the lens, the cortex, but not in the lens nucleus where older proteins are present. The 47 kDa component was composed of betaB1-, betaB2- and gammaS-crystallin, with the gammaS-crystallin having glutathione bound at Cys-82 and at Cys-22, Cys-24 or Cys-26. We conclude that when glutathione becomes bound to gammaS-crystallin, it causes it to bind in turn to the beta-crystallin polypeptides to form a dimer.
 ...
PMID:The identification of a reaction site of glutathione mixed-disulphide formation on gammaS-crystallin in human lens. 1476 3

Chromium is released during several industrial processes and has accumulated in some estuarine areas. Its effects on mammals have been widely studied, but relatively little information is available on its effects on fish. Gene expression changes are useful biomarkers that can provide information about toxicant exposure and effects, as well as the health of an organism and its ability to adapt to its surroundings. Therefore, we investigated the effects of Cr(VI) on gene expression in the sediment dwelling fish, winter flounder (Pseudopleuronectes americanus). Winter flounder ranging from 300 to 360 g were injected i.p. with Cr(VI) as chromium oxide at 25 microg/kg chromium in 0.15N KCl. Twenty-four hours following injections, winter flounder were euthanized with MS-222 and the livers were excised. Half of the livers were used to make cytosol and the other half were used to isolate mRNA for subtractive hybridization. Subtractive clones obtained were spotted onto nylon filters, which revealed several genes with potentially altered expression due to Cr(VI), including an alpha class GST, 1-Cys peroxiredoxin (a non-selenium glutathione peroxidase), a P-450 2X subfamily member, two elongation factors (EF-1 gamma and EF-2), and complement component C3. Semi-quantitative RT-PCR was performed and confirmed that Cr(VI) down-regulated complement component C3, an EST, and two potential glutathione peroxidases, GSTA3 and 1-Cys peroxiredoxin. In addition, cytosolic GSH peroxidase activity was reduced, and silver stained SDS-PAGE gels from glutathione-affinity purified cytosol demonstrated that a 27.1 kDa GSH-binding protein was down-regulated greater than 50%. Taken together, Cr(VI) significantly altered the expression of several genes including two potential glutathione peroxidases in winter flounder.
 ...
PMID:Construction of a subtractive library from hexavalent chromium treated winter flounder (Pseudopleuronectes americanus) reveals alterations in non-selenium glutathione peroxidases. 1500 2

Oxidation of the food preservative 2,6-di-tert-butyl-4-methylphenol (BHT) by mouse lung cytochrome P450 produces electrophilic quinone methides thought to promote lung tumors in mice by covalent binding to critical proteins. Specific pulmonary targets of 2,6-di-tert-butyl-4-methylenecyclohexa-2,5-dienone (BHT-QM) have not been identified, however. The present work was undertaken to determine if glutathione S-transferase P1-1 (GSTP1-1) is alkylated by BHT-QM, as this protein is overexpressed in tumors and has important roles in protecting cells from electrophiles and oxidants and in regulating stress kinases. This work was conducted with cell lines C10 and E10 derived from mouse lung epithelia and their spontaneous transformants, the tumorigenic cell lines A5 and E9. Cytosolic GSTs were isolated by affinity chromatography and analyzed by ESI-LC/MS. Ion current chromatograms indicated that GSTP1 predominates over the other isoforms, especially in tumorigenic cells. Treatment with BHT-QM inhibited cytosolic GST activity by 28-44%, and inhibition was exacerbated by depleting intracellular GSH. Alkylation of GSTP1 by BHT-QM was investigated by separating cytosolic proteins with two-dimensional SDS-PAGE and detecting adducts by Western blotting with polyclonal antibodies that recognize the BHT group. The identity of GSTP1 comigrating with immunoreactive material was confirmed by in-gel proteolysis and LC/MS/MS analysis. Human GSTP1 was utilized to investigate the specific residues involved in QM binding. The only peptide adduct detected in digests of monoadducted GSTP1 corresponded to Cys101, whereas adducts at Cys14, Cys47, and Cys101 were identified from the trialkylated protein. Losses of transferase activity were most influenced by alkylation at Cys47, but binding to Cys14 appeared to inhibit the activity further. These findings demonstrate that cytosolic GSTP1 may be a target for BHT-QM resulting in decreased cellular protection from electrophiles and oxidants. Alkylation also may interfere with GSTP1 regulation of stress kinases, thereby influencing phosphorylation and cell growth.
 ...
PMID:Inhibition of glutathione S-transferase P1-1 in mouse lung epithelial cells by the tumor promoter 2,6-di-tert-butyl-4-methylene-2,5-cyclohexadienone (BHT-quinone methide): protein adducts investigated by electrospray mass spectrometry. 1560 44

Evidence suggests that aging, per se, is a major risk factor for cardiac dysfunction. Oxidative modification of cardiac proteins by non-enzymatic glycation, i.e. advanced glycation endproducts (AGEs), has been implicated as a causal factor in the aging process. This study was designed to examine the role of aging on cardiomyocyte contractile function, cardiac protein oxidation and oxidative modification. Mechanical properties were evaluated in ventricular myocytes from young (2-month) and aged (24-26-month) mice using a MyoCam system. The mechanical indices evaluated were peak shortening (PS), time-to-PS (TPS), time-to-90% relengthening (TR90) and maximal velocity of shortening/relengthening (+/- dL/dt). Oxidative stress and protein damage were evaluated by glutathione and glutathione disulfide (GSH/GSSG) ratio and protein carbonyl content, respectively. Activation of NAD(P)H oxidase was determined by immunoblotting. Aged myocytes displayed a larger cell cross-sectional area, prolonged TR90, and normal PS, +/- dL/dt and TPS compared with young myocytes. Aged myocytes were less tolerant of high stimulus frequency (from 0.1 to 5 Hz) compared with young myocytes. Oxidative stress and protein oxidative damage were both elevated in the aging group associated with significantly enhanced p47phox but not gp91phox expression. In addition, level of cardiac AGEs was approximately 2.5-fold higher in aged hearts than young ones determined by AGEs-ELISA. A group of proteins with a molecular range between 50 and 75 kDa with pI of 4-7 was distinctively modified in aged heart using one- or two-dimension SDS gel electrophoresis analysis. These data demonstrate cardiac diastolic dysfunction and reduced stress tolerance in aged cardiac myocytes, which may be associated with enhanced cardiac oxidative damage, level of AGEs and protein modification by AGEs.
 ...
PMID:Aging induces cardiac diastolic dysfunction, oxidative stress, accumulation of advanced glycation endproducts and protein modification. 1577 9

A glutathione S-transferase (GST) from Lactuca sativa was purified to electrophoretic homogeneity approximately 403-fold with a 9.6% activity yield by DEAE-Sephacel and glutathione (GSH)-Sepharose column chromatography. The molecular weight of the enzyme was determined to be approximately 23,000 by SDS-polyacrylamide gel electrophoresis and 48,000 by gel chromatography, indicating a homodimeric structure. The activity of the enzyme was significantly inhibited by ShexylGSH and S-(2,4-dinitrophenyl) glutathione. The enzyme displayed activity towards 1-chloro-2,4-dinitrobenzene, a general GST substrate and high activities towards ethacrynic acid. It also exhibited glutathione peroxidase activity toward cumene hydroperoxide.
 ...
PMID:Purification and biochemical properties of glutathione S-transferase from Lactuca sativa. 1582 2

The present investigation is aimed to identify and characterize tumor-associated antigen (TAA) in animals exposed to hepatocarcinogen. Swiss albino mice showed an enhanced expression of an approximately 58 kDa glycoprotein in liver cells upon exposure to a potential hepatocarcinogen N-nitrosodibutylamine (DBN). Carcinogenesis induction in mice was monitored by assays of y-glutamyl transpeptidase (GGT), acetylcholine esterase (AChE), glutathione-S-transferase (GST) activities and the level of glutathione (GSH) in liver. DBN treated animals showed cell distortion and extensive necrosis as observed by histological examination. The over-expressed TAA was purified by ion-exchange chromatography and further characterized by SDS-PAGE. The carbohydrate contents and glycan linkage to the polypeptide backbone was analyzed by using the DIG glycan differentiation and de-glycosylation kits. The glycoprotein has glycan chains that are N-linked via asparagines to the polypeptide backbone. It was also observed that the molecule is rich in sialic acid residues with a significantly high carbohydrate to protein ratio (> 2:1). The over-expressed high molecular weight glycoprotein TAA was found to be highly immunogenic and could eventually be used to induce immune response in order to counter tumor regression.
 ...
PMID:Molecular characterization of tumor associated antigen in mice exposed to a hepatocarcinogen. 1588 69

Protein-glutathione mixed disulfide formation was investigated in vitro by exposure of human platelets to the thiol-specific oxidant azodicarboxylic acid-bis-dimethylamide (diamide). We found that diamide causes a decrease in the reduced form of glutathione (GSH), paralleled by an increase in protein-GSH mixed disulfides (S-glutathionylated proteins), which was not accompanied by any significant increase in the basal level of glutathione disulfide (GSSG). The increase in the appearance of S-glutathionylated proteins was inversely correlated with ADP-induced platelet aggregation. Platelet cytoskeleton was analyzed by SDS-PAGE followed by Western immunoblotting with anti-GSH antibody. The main S-glutathionylated cytoskeletal protein proved to be actin, which accounts for 35% of the platelet total protein content. Our results suggest that neither GSSG formation nor a consequent thiol-disulfide exchange mechanism is involved in actin S-glutathionylation of human platelets exposed to diamide. Instead, a mechanism involving the initial oxidative activation of actin thiol groups, which then react with GSH to the protein-GSH mixed disulfides, makes it likely that platelet actin is S-glutathionylated without any significant increase in the GSSG content.
 ...
PMID:S-glutathionylation in human platelets by a thiol-disulfide exchange-independent mechanism. 1589 Jun 24


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>