A Comparative Study of Metallothionein Gene Expression in Peripheral Lymphocytes and Blood Cadmium Level among Die Casting Male Workers
The Zrt/Irt-related protein 8 (ZIP8) encoded by slc39a8 is now emerging as an important zinc transporter involved in cellular cadmium incorporation. We have previously shown that mRNA and protein levels of ZIP8 were decreased in cadmium-resistant metallothionein-null (A7) cells, leading to a decreas …
These studies were designed to investigate the effects of low dietary zinc, copper or selenium intake of certain types of stress and of injection of zinc, copper or cadmium on metallothionein I (MT-I) concentrations in the blood and urine of rats. The aim was to establish whether such measurements could be of value in the diagnosis of zinc deficiency. Marginal zinc deficiency rapidly caused a major decrease in MT-I levels in the blood cells and to a lesser extent in urine. Injection of zinc and also of cadmium and copper had the opposite effect and increased MT-I concentrations in both samples, although the effects of zinc on blood cells and urine were relatively transient. The MT-I in the blood cells was associated mainly with the erythrocytes. No changes in blood or urine MT-I levels were found in copper- and selenium-deficient rats. Neither cold stress nor restriction of food intake for 24 h had any significant effect on MT-I levels in the blood cells or urine. Endotoxin injection increased urinary
OBJECTIVES: We examined the distribution of metallothionein (MT), a stress-inducible protein, and the cardiomyocyte diameter in human hearts after left-ventricular assist device (LVAD) support. BACKGROUND: Remodeling in end-stage heart failure is cha
The overall strategy is to employ a well-characterized soil organism to probe the regulation of a key gene in metal detoxification and homeostasis. The aim of the research is to identify metal regulatory elements (MREs) within the promoters of two C. elegans MT genes, and isolate and characterize proteins that interact with the candidate MREs. Candidate MREs will be identified by site-directed mutagenesis and deletion experiments. The functional significance of each modification will be determined in vivo (inside the organism) by creating transgenic nematodes containing a ß-galactosidase reporter gene. Changes in the level of pattern of reporter gene activity in cadmium-exposed nematodes will be used to identity functional promoter elements. Metal-dependent interaction of crude extract proteins with candidate MREs will be evaluated by gel mobility shift assay and DNaseI footprinting. Regulatory proteins that interact with candidate MREs will be isolated from extracts prepared from control and ...
We studied how the protein metallothionein (MT) impacts arsenic-induced oxidative DNA damage (ODD) using cells that poorly express MT (MT-I/II double knockout embryonic cells; called MT-null cells) and wild-type (WT) MT proficient cells. protein) and in the metal-responsive transcription element-1 (MTF-1) which is required to induce the MT gene. In contrast basal MT levels were… Continue reading We studied how the protein metallothionein (MT) impacts arsenic-induced oxidative DNA. ...
In this study, we demonstrate that the high-affinity component of the manganese transport system in mammalian cells is also used for cadmium uptake. Previously, we have established cadmium-resistant cell lines from MT-null mouse cells that exhibited a marked decrease in the uptake of cadmium (Yanagiya et al., 1999). The application of multitracer technique in this study revealed that the uptake of ultra-trace amount of manganese in Cd-rB5 cells was reduced to approximately 10% of that in parental cells, whereas no change in the incorporation of zinc, copper, or iron was observed. Because cadmium accumulation in the Cd-rB5 cells was also reduced to 10% of that in parental cells, the same mechanism may be responsible for the reduction of the incorporation of both cadmium and manganese into Cd-rB5 cells.. The results of time- and dose-dependent uptake of Mn2+ in Cd-rB5 and parental cells (Fig. 2, A and C) suggest that there are at least two components, having high and low affinity to Mn2+, for ...
Title: Role of Metallothionein in Inflammatory Lung Diseases. VOLUME: 5 ISSUE: 1. Author(s):Ken-ichiro Inoue, Hirohisa Takano, Akinori Shimada and Masahiko Satoh. Affiliation:Environmental Health Sciences Division, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba 305-8506, Japan.. Keywords:Metallothionein, lung inflammation, lipopolysaccharide, allergen, ozone. Abstract: The integration of knowledge concerning the regulation of metallothionein (MT) with research on its proposed functions is necessary to clarify how MT affects cellular processes. MT expression is induced/enhanced in various tissues by a number of physiological mediators through several response elements in the MT gene promoter. The cellular accumulation of MT depends on the availability of cellular zinc derived from the diet. MT modulates: 1) the binding and exchange/ transport of heavy metals such as zinc, cadmium, or copper under physiological conditions and cytoprotection from their toxicities, and 2) the ...
1. Introduction. The necessity of large scale supply of animal protein for a growing population simultaneously to a sharp decrease in natural fisheries occurring during the past two decades has resulted in a worldwide spread of aquaculture of many aquatic species. Shrimps are one of the most successful aquaculture products with a well established technology for large scale intensive and semi-intensive cultivation. The high productivity achieved in shrimp aquaculture is dependent on a large supply of aquafeeds (5 to 10 t.ha-1.year-1) with a resultant productivity of 2 to 6 t.ha-1.year-1. Chemical composition of aquafeeds is variable, concentrations of major nutrients, protein and fat contents are relatively controlled. However, minor components, including trace metals mostly originally present in aquafeeds as natural constituents of the original raw materials, are not controlled (Boyd and Massaut, 1999; Tacon and Forster, 2003; Usydus et al., 2009). For example, concentrations of Cd in shrimp ...
Zinc is essential for many cellular processes, including DNA synthesis, transcription, and translation, but excess can be toxic. A zinc-induced gene, smtA, is required for normal zinc-tolerance in the cyanobacterium Synechococcus PCC 7942. Here we report that the protein SmtA contains a cleft lined with Cys-sulfur and His-imidazole ligands that binds four zinc ions in a Zn4Cys9His2 cluster. The thiolate sulfurs of five Cys ligands provide bridges between the two ZnCys4 and two ZnCys3His sites, giving two fused six-membered rings with distorted boat conformations. The inorganic core strongly resembles the Zn4Cys11 cluster of mammalian metallothionein, despite different amino acid sequences, a different linear order of the ligands, and presence of histidine ligands. Also, SmtA contains elements of secondary structure not found in metallothioneins. One of the two Cys4-coordinated zinc ions in SmtA readily exchanges with exogenous metal (111Cd), whereas the other is inert. The thiolate sulfur ...
A mouse cDNA clone, M96, encoding a metal-regulating-element (MRE)-binding protein, was analysed for its ability to act as a metal-regulated transcription factor. The metal depletion of a glutathione S-transferase (GST)-M96 fusion protein showed that Zn2+ ions modulate the MRE-binding activity, suggesting that the M96-encoded protein is a Zn2+-regulated factor (ZiRF1). The methylation interference assay showed the specific interactions of ZiRF1 with the MRE, MREd/c, present on the mouse metallothionein Ia promoter. Point mutations of the MREd/c nullified the metal-regulatory properties of this region. In mouse L-cell nuclear extracts, mobility-shift assays revealed a Zn2+-dependent MRE-binding complex (MBC) with DNA-recognition properties similar to those of ZiRF1. Antibodies raised against purified GST-ZiRF1 were able to specifically recognize MBC in Western-blot analyses. Competition analysis of MRE-binding proteins from mouse NIH3T3 cells with oligonucleotide matching the binding sites for ...
Many diabetic patients suffer from cardiomyopathy, even in the absence of vascular disease. This diabetic cardiomyopathy predisposes patients to heart failure and mortality from myocardial infarction. Evidence from animal models suggests that reactive oxygen species play an important role in the development of diabetic cardiomyopathy. Our laboratory previously developed a transgenic mouse model with targeted overexpression of the antioxidant protein metallothionein (MT) in the heart. In this study we used MT-transgenic mice to test whether an antioxidant protein can reduce cardiomyopathy in the OVE26 transgenic model of diabetes. OVE26 diabetic mice exhibited cardiomyopathy characterized by significantly altered mRNA expression, clear morphological abnormalities, and reduced contractility under ischemic conditions. Diabetic hearts appeared to be under oxidative stress because they had significantly elevated oxidized glutathione (GSSG). Diabetic mice with elevated cardiac MT (called OVE26MT mice) ...
Janssen, H. H. and Dallinger, R. (1991): Diversification of cadmium-binding proteins due to different levels of contamination in Arion lusitanicus , Arch Environ Contam Toxicol ...
The dynamics of metal binding to and transfer from metalloproteins involved in metal homeostasis are important for understanding cellular distribution of metal ions. The dicotyledonous plant Arabidopsis thaliana has two type 4 seed-specific metallothionein homologues, MT4a and MT4b, with likely roles in zinc(II) homeostasis. These two metallothioneins are 84% identical, with full conservation of all metal-binding cysteine and histidine residues. Yet, differences in their spatial and temporal expression patterns suggested divergence in their biological roles. To investigate whether biological functions are reflected in molecular properties, we compare aspects of zinc(II)-binding dynamics of full-length MT4a and MT4b, namely the pH dependence of zinc(II) binding and protein folding, and zinc(II) transfer to the chelator EDTA. UV-Vis and NMR spectroscopies as well as native electrospray ionisation mass spectrometry consistently showed that transfer from Zn6MT4a is considerably faster than from ...
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Full Text CA-92-22 POSSIBLE ROLE OF METALLOTHIONEIN IN CARCINOGENESIS NIH GUIDE, Volume 21, Number 32, September 4, 1992 RFA: CA-92-22 P.T. 34 Keywords: Cancer/Carcinogenesis Toxicology Biochemistry, Proteins Biology, Molecular Biomedical Research, Multidiscipl National Cancer Institute Letter of Intent Receipt Date: October 2, 1992 Application Receipt Date: December 8, 1992 PURPOSE The Chemical and Physical Carcinogenesis Branch, Division of Cancer Etiology (DCE), National Cancer Institute (NCI), in collaboration with the Cancer Biology Branch, Division of Cancer Biology, Diagnosis, and Centers (DCBDC), NCI, and the Grants and Contracts Operations Branch, Division of Cancer Treatment (DCT), NCI, invites investigator-initiated research grant applications to elucidate the possible role of metallothionein (MT) in carcinogenesis. New and experienced investigators in relevant fields and disciplines may apply for funds to pursue this research. Historically, three major groups of scientists have ...
Complementary DNA encoding a facilitative glucose transporter was isolated from a human hepatoma cell line (HepG2) cDNA library and subcloned into a metal-inducible mammalian expression vector, pLEN (California Biotechnology) containing human metallothionein gene II promoter sequences. Chinese hamster ovary (CHO) cells transfected with this transporter expression vector, pLENGT, exhibited a 2-17-fold increase in immunoreactive HepG2-type glucose transporter protein, as measured by protein immunoblotting with antipeptide antibodies directed against the HepG2-type glucose transporter C-terminal domain. Expression of the human glucose transporter was verified by protein immunoblotting with a mouse polyclonal antiserum that recognizes the human but not the rodent HepG2-type transporter. 2-Deoxy-D-glucose uptake was increased 2-7-fold in transfected cell lines. Polyclonal antisera directed against purified red blood cell glucose transporter were raised in several rabbits. Antiserum from one rabbit, delta,
Annealing Product Cycle Accession Gene Abreviation Primer sequence temperature length number number ------------------ ------------- ------------------------- ----------- ------- ------ --------- Hsp27-1 Hsp27-1 CGCGCTCAGCCGGCAACTCAG 64 419 27 XM_055937 Hsp27-1 Hsp27-1 AGGGGTGGGCATCCAGGCTAAGG 64 419 27 XM_055937 Hsp27-2 Hsp27-2 TCCTGACCCCCACACTCTACCA 61 421 27 NM_001541 Hsp27-2 Hsp27-2 GCTGCCTCCTCCTCTTCCTCTG 61 421 27 NM_001541 aA-crystallin aA CCACCTCGGCTCCCTCGTCCTAAG 64 492 25 NM_000394 aA-crystallin aA CCATGTCCCCAAGAGCGGCACTAC 64 492 25 NM_000394 RPL13a RPL13a GTATGCTGCCCCACAAAACCA 58 387 25 XM_027885 RPL13a RPL13a CAACGCATGAGGAATTAACAGTCTT 58 387 25 XM_027885 Metallothionein IF MTIF GCTTCTCTCTTGGAAAGTCC 55 226 30 M10943 Metallothionein IF MTIF GGCATCAGTCGCAGCAGCTG 55 226 30 M10943 Metallothionein IH MTIH GAACTCCAGTCTCACCTCGG 55 213 30 X64834 Metallothionein IH MTIH GACATCAGGCACAGCAGCTG 55 213 30 X64834 Metallothionein IG MTIG GCCTCTTCCCTTCTCGCTTG 55 234 30 XM_048213 Metallothionein IG MTIG ...
The earliest perception restricted the occurrence of metallothioneins to animals, fungi, and certain bacteria and assigned the corresponding functions in plants to the enzymatically synthesized phytochelatins. This picture has now clearly changed, and the existence of plant metallothioneins is generally accepted. Compared to the vertebrate forms, plant metallothioneins show an intriguing sequence diversity, which is contrasted by an enormous lack of information about their possible functions, properties, and structures. The majority of functions proposed is derived from gene expression studies, which are reviewed with a not exhaustive but representative list of examples. However, it becomes evident that so far it is not possible to draw a clear-cut picture that ascribes specific functions to a given metallothionein. Rather, proposed functions overlap or are complementary to each other; sometimes even contradictory results are obtained. Regarding the structures and properties of the plant ...
Metallothionein (MT) is a family of cysteine-rich, low molecular weight (MW ranging from 500 to 14000 Da) proteins. They are localized to the membrane of the Golgi apparatus. MTs have the capacity to bind both physiological (such as zinc, copper, selenium) and xenobiotic (such as cadmium, mercury, silver, arsenic) heavy metals through the thiol group of its cysteine residues, which represent nearly 30% of its constituent amino acid residues. MT was discovered in 1957 by Vallee and Margoshe from purification of a Cd-binding protein from horse (equine) renal cortex. MTs function is not clear, but experimental data suggest MTs may provide protection against metal toxicity, be involved in zinc and copper regulation, and provide protection against oxidative stress. There are four main isoforms expressed in humans (family 1, see chart below): MT1 (subtypes A, B, E, F, G, H, L, M, X), MT2, MT3, and MT4. In the human body, large quantities are synthesised primarily in the liver and kidneys. Their ...
Malignant tumours of head and neck localisation represent serious health problem. All over the world, there is about 650 000 of patients who fall ill with head and neck carcinoma per year. Treatment of this malignity is based especially on the well-timed surgery intervention and radiotherapy. Therefore, alternative ways of diagnostics, which would replace commonly used biopsy in combination with computed tomography, are intensely searched. Attention is devoted especially to molecules, whose levels are changed in body due to malignant process and malignant disease progression. The main objective of this study consisted in determination of metallothionein (/MT)/ levels in tumour tissues of patients suffering from head and neck tumours using differential pulse voltammetry Brdicka reaction. Fifty-five samples of tumour tissue (45 % laryngeal, 40 % oropharyngeal, 9 % hypopharyngeal and 6 % oral cavity cancer) were included in this study. After describing of differences in differential pulse ...
Günther, V; Lindert, U; Schaffner, W (2012). The taste of heavy metals: Gene regulation by MTF-1. Biochimica et Biophysica Acta, 1823(9):1416-1425.. Günther, V; Strassen, T; Lindert, U; Dagani, P; Waldvogel, D; Georgiev, O; Schaffner, W; Bethge, T (2012). Simian virus 40 (SV40) strains with novel properties generated by replacing the viral enhancer with synthetic oligonucleotides. Journal of Virology, 86(6):3135-3142.. Lindert, U; Cramer, M; Meuli, M; Georgiev, O; Schaffner, W (2009). Metal-responsive transcription factor 1 (MTF-1) activity is regulated by a nonconventional nuclear localization signal and a metal-responsive transactivation domain. Molecular and Cellular Biology, 29(23):6283-6293.. Lindert, U. Comparison and functional characterization of the metal-responsive transcription factor 1 (MTF-1) from rodents and human. 2009, University of Zurich, Faculty of Science.. Lindert, U; Leuzinger, L; Steiner, K; Georgiev, O; Schaffner, W (2008). Characterization of metal-responsive ...
Metallothioneins (MTs) are members of a family of cysteine-rich low molecular weight polypeptides which play an important role in heavy metal detoxification and homeostasis of intracellular metal ions
A rapid and sensitive one-vial procedure to determine metallothionein (MT) containing zinc (Zn) and cadmium (Cd) is described. New features of this Cd-saturation method are: high molecular weight...
The first cyclic analog of a metallothionein (MT) was prepared and analyzed by UV and (magnetic) circular dichroism spectroscopy, ESI-MS as well as NMR spectroscopy. Results reveal that the evaluated cyclic g-Ec-1 domain of the wheat MT Ec-1 retains its ability to coordinate two Zn(II) or Cd(II) ions and adopts a three-dimensional structure that is highly similar to the one of the linear wild-type form. However, the reduced flexibility of the protein backbone facilitates structure solution significantly and results in a certain stabilization of metal binding to the protein.
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Genes encoding the small, metal-binding metallothioneins (MTs) are found in all eukaryotes (often in multiple copies) as well as some prokaryotes (1). MTs are unusually rich in cysteine residues that coordinate multiple zinc and copper atoms under physiological conditions. In the mouse, there are four MT genes that reside in a 50-kb region on chromosome 8, whereas humans have at least 16 MT genes clustered on chromosome 16 (2, 3). The mouse MT I and II genes are expressed at all stages of development in many cell types of most organs; they are coordinately regulated by metals, glucocorticoids, and inflammatory stress signals (4). MT III is expressed predominantly in neurons but also in glia and male reproductive organs (5-7). MT IV is expressed in differentiating stratified squamous epithelial cells (3). All four MT genes are expressed in the maternal deciduum (8). The fact that there are multiple MT genes, expressed in distinct patterns, suggests that they should have important functions; ...
I dont treat rheumatoid arthritis directly as my work focuses on brain related disorder, though there can be a great deal of overlap with autoimmune issues. Part of the reason may have to do with the micro biome (I recently wrote a post on that) and secondary gut permeability and the significant role of zinc in the immune system, the digestive system and the brain functioning. A related by deeper connection maybe the metallothionein proteins (which are dependent on zinc to function). MT is a major (though relatively unknown) antioxidant that controls levels of zinc and copper. But it also relates to the functioning of the intestinal and blood brain barriers, protection of enzymes that break down casein and gluten and causes a decrease in inflammation after injury and illness among many other things. As you can see, if it is not working well due to zinc deficiency for example, or because of a genetic issue, people can have copper overload as well as a range of other issues. From a clinical ...
A cDNA for plaice metallothionein (MT) was cloned and the sequence for the first 45 amino acid residues was deduced. The sequence of plaice MT was exactly homologous to that of winter flounder MT. Studies of MT induction by Cd and Zn in a rainbow trout (RTH) cell line showed that both metals were strong inducers of the protein in contrast to previous reports which have shown little induction of MT mRNA by Cd. Translational control or differential mRNA and MT stabilities are therefore inferred. ...
The goals of this study were to determine the expression of metallothionein isoform 1 and 2 proteins (MT-1 and MT-2) in bladder cancer and then to determine which MT isoform-specific genes promoted the expression of these proteins ...
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Description: Metallothionein SmtA; Contain 9 cysteine residues; plays a role in essential metal ion homeostasis (especially zinc homeostasis) and resistance to certain non-essential metal ions. Metallothioneins have a high content of cysteine residues that bind various heavy metals. Cells that lack of metallothionein have reduced uptake of Zn++ and reduced tolerance to high Zn++ concentrations (althogh the Cu++ and Hg++ tolerances are unchanged); often Cd++ resistant when cells grow in high level of Cd++ ...
Metallothioneins have a high content of cysteine residues that bind various heavy metals; these proteins are transcriptionally regulated by both heavy metals and glucocorticoids.
ABSTRACT: The body concentrations of Ag in a top marine predator, the whelk Thais clavigera, varied significantly (1.10 to 4.67 µg g-1 dry wt) in individuals collected from 3 intertidal rocky shores in Hong Kong, whereas there was no significant difference in Ag concentrations in species from lower trophic levels (macroalgae Ulva sp. and rocky oyster Saccostrea cucullata). We compared the variation in Ag biodynamics in whelks collected from these 3 locations. There was no significant difference in the Ag dietary assimilation efficiency, Ag dissolved uptake rate, or estimated metallothionein concentration among the 3 locations. Significant differences were found in the Ag efflux rate constant and the subcellular distribution of Ag. Whelks from Clear Water Bay, which had the lowest body concentration of Ag, had a higher Ag efflux rate constant (0.0268 d-1) than those collected from the other 2 sites (0.0076 to 0.0095 d-1), as well as a correspondingly lower fraction of Ag in the ...
Metallothioneins have a high content of cysteine residues that bind various heavy metals; these proteins are transcriptionally regulated by both heavy metals and glucocorticoids.
Binds heavy metals. Contains three zinc and three copper atoms per polypeptide chain and only a negligible amount of cadmium. Inhibits survival and neurite formation of cortical neurons in vitro ...
properties of metal binding and free radical scavenging to the intricate workings of brain. In this compilation of four reports, first delivered at the 11th International Neurotoxicology Association (INA-11) Meeting, June 2007, the authors present the work of their laboratories, each of which gives an important insight into the actions of MT in the brain. What emerges is that MT has the potential to contribute to a variety of processes, including neuroprotection, regeneration, and even cognitive functions. In this article, the properties and CNS expression ofMTare briefly reviewed before Dr Hidalgo describes his pioneering work using transgenic models of MTexpression to demonstrate how this protein plays a major role in the defence of the CNS against neurodegenerative disorders and other CNS injuries. His groups work leads to two further questions, what are the mechanisms at the cellular level by which MTacts, and does this protein influence higher order issues of architecture and cognition. ...
TY - JOUR. T1 - Transgelin-2 as a therapeutic target for asthmatic pulmonary resistance. AU - Yin, Lei Miao. AU - Xu, Yu Dong. AU - Peng, Ling Ling. AU - Duan, Ting Ting. AU - Liu, Jia Yuan. AU - Xu, Zhijian. AU - Wang, Wen Qian. AU - Guan, Nan. AU - Han, Xiao Jie. AU - Li, Hai Yan. AU - Pang, Yu. AU - Wang, Yu. AU - Chen, Zhaoqiang. AU - Zhu, Weiliang. AU - Deng, Linhong. AU - Wu, Ying Li. AU - Ge, Guang Bo. AU - Huang, Shuang. AU - Ulloa, Luis. AU - Yang, Yong Qing. PY - 2018/2/7. Y1 - 2018/2/7. N2 - There is a clinical need for new bronchodilator drugs in asthma, because more than half of asthmatic patients do not receive adequate control with current available treatments. We report that inhibition of metallothionein-2 protein expression in lung tissues causes the increase of pulmonary resistance. Conversely, metallothionein-2 protein is more effective than -2-agonists in reducing pulmonary resistance in rodent asthma models, alleviating tension in tracheal spirals, and relaxing airway smooth ...
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These reference sequences exist independently of genome builds. Explain. These reference sequences are curated independently of the genome annotation cycle, so their versions may not match the RefSeq versions in the current genome build. Identify version mismatches by comparing the version of the RefSeq in this section to the one reported in Genomic regions, transcripts, and products above. ...
As part of the International Genetically Engineered Machine (iGEM) competition, our team is investigating new methods of increasing the efficiency of bioremediation and biosynthesis using modified bacteria flagellin. By making chimeric insertions in the variable domain of the flagellin, we can incorporate metal binding proteins, enzymes, adhesive proteins as well as scaffolding proteins to further extend the possible applications.
As part of the International Genetically Engineered Machine (iGEM) competition, our team is investigating new methods of increasing the efficiency of bioremediation and biosynthesis using modified bacteria flagellin. By making chimeric insertions in the variable domain of the flagellin, we can incorporate metal binding proteins, enzymes, adhesive proteins as well as scaffolding proteins to further extend the possible applications.
A variety of cellular proteins bind to cellular and viral enhancer elements. One such factor, known as AP-2, is a 52-kDa transcription factor identified by its interaction with the SV40 and metallothionein enhancers. In addition, it has been found that AP-2 binds to the SV40 T-antigen. AP-2 activity …
Complete information for MT1M gene (Protein Coding), Metallothionein 1M, including: function, proteins, disorders, pathways, orthologs, and expression. GeneCards - The Human Gene Compendium
In article ,1995May15.212901.16374 at rs6000.cmp.ilstu.edu,, ajotsuka at rs6000.cmp.ilstu.edu (Anthony Otsuka) wrote: , , We have been using pRSET vectors (Invitrogen) for expressing , proteins. During purification over nickel chelating columns, , we see a considerable amount of background protein binding , (other metal-binding proteins?). Has anyone been able to , achieve ,90% purity by using the single column? Any advice , would be appreciated. Thanks, A. Otsuka Hi, I«ve been working for now more than 2 years on expressing some human ribosomalproteins using the pRSET vectors (Expression is done in BL21 or BLR from Novagen). Because my proteins are expressed in inclusion-bodies, i«m doing the purification under denaturing conditions followed by a renaturation step. Usually I get more than 95% purity on a single column. Sometimes, especially on purifying whole bacteria lysates on the resin, my preparations contain a impurity, which appears at about 29 kD in SDS-PAGE. I can find this impurity ...
CP000529.PE173 Location/Qualifiers FT CDS_pept 186561..187079 FT /codon_start=1 FT /transl_table=11 FT /locus_tag="Pnap_0175" FT /product="rod shape-determining protein MreD" FT /note="PFAM: Rod shape-determining protein MreD; KEGG: FT pol:Bpro_0224 putative rod shape-determining MreD FT transmembrane protein" FT /db_xref="EnsemblGenomes-Gn:Pnap_0175" FT /db_xref="EnsemblGenomes-Tr:ABM35500" FT /db_xref="GOA:A1VIM2" FT /db_xref="InterPro:IPR007227" FT /db_xref="InterPro:IPR026034" FT /db_xref="UniProtKB/TrEMBL:A1VIM2" FT /protein_id="ABM35500.1" FT /translation="MIMRSGQQLLLPASPVFIWCTLIAALLLDMLPLGRIPWMPEFLAL FT VLVFWSVHQPLKVGIGIAFMFGLAIDVHQTSLLGQHAFSYTMLSFLAAMIQRRLLWFTV FT PLQALQVLPLFAAAHGAEIILRLLGGGVFPGWIVLLAPLAEALLWPVASVLLLVPQLRT FT PDRDENRPL" atgatcatgc gttccggcca gcaactgctg ctgcccgcca gtccggtgtt catctggtgc 60 accctgattg cggccttgct gctcgacatg ctgccgctgg gacgcatccc ctggatgccc 120 gaatttctgg cgctggtgct ggtgttctgg agcgtgcacc agccgctcaa agtcggcatc 180 ggcattgcct tcatgtttgg cttggcgata gatgtgcacc ...
Drought stress is one of the serious problems that restricted agronomic plant production worldwide. In molecular level, the harmful effect of drought stress is mostly caused by producing of large amount of reactive oxygen species (ROS). Catalase and Metallothionein genes have a crucial role to mope the hydrogen peroxide (H2O2) resulting reducing oxidative damage. In this research the gene expression pattern of Catalase and Metallothionein was studied in response to drought stress treatments. The treatments included - 0.3 bar, - 0.9 bar, - 8 bar and -12 bar and wheat varieties included Zagros (drought tolerant), Moghan (semi- tolerant) and Tajan (drought sensitive). The amount of cellular oxidative levels (TBARM) increased steady by intensify of drought stress levels. Real time PCR analysis showed different expression pattern for catalase and metallothionein encoded genes. Catalase gene expression was increased during drought stress up to -8 bar and reduced in -12 bar treatment, in all cultivars
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In contrast to many other genes containing a CpG island, the testis-specific H2B (TH2B) histone gene exhibits tissue-specific methylation patterns in correlation with gene activity. Characterization of the methylation patterns within a 20-kb segment containing the TH2A and TH2B genes in comparison with that in a somatic histone cluster revealed that: (i) the germ cell-specific unmethylated domain of the TH2A and TH2B genes is defined as a small region surrounding the CpG islands of the TH2A and TH2B genes and (ii) somatic histone genes are unmethylated in both liver and germ cells, like other genes containing CpG islands, whereas flanking sequences are methylated. Transfection of in vitro-methylated TH2B, somatic H2B, and mouse metallothionein I constructs into F9 embryonal carcinoma cells revealed that the CpG islands of the TH2A and TH2B genes were demethylated like those of the somatic H2A and H2B genes and the metallothionein I gene. The demethylation of those CpG islands became ...
Expression of intracellular metallothionein (MT) has been linked to cis-diamminedichloroplatinum (cDDP) resistance in human germ cell tumor cell lines. To determine whether exposure to cDDP would select for cells with increased MT expression, the MT content of the human teratocarcinoma cell line T7800 was measured after development of resistance to cDDP by exposure to progressively higher drug concentrations (6.25-25 microM). cDDP-resistant cells (T7800R) had significantly higher MT mRNA and MT protein, increased resistance to killing by cDDP and altered in vitro growth kinetics compared to parental T7800 cells. cDDP resistance in a variety of other human tumor cell lines correlated with MT content, with no significant difference in glutathione level. These data indicate that selection in vitro for cDDP resistance in human germ cell tumors coselects for cells with enhanced MT content. However, selected cells differed in characteristics other than MT content. They had a slower growth rate and, ...