• Bulletin of the Korean Chemical Society
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• v.35 no.4
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• pp.1087-1090
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• 2014

A simple and general method that selectively introduces metal binding sites into a protein can greatly increase the ability to design and biosynthesize artificial metalloproteins. Here, we report the incorporation of a phenanthroline-containing amino acid (Phen-Ala) into proteins in Escherichia coli by using the $tRNA{^{Tyr}}_{CUA}$ and tyrosyl aminoacyl-tRNA synthetase pair (BpyRS) from Methanococcus jannaschii, which was originally developed for a bipyridine-containing amino acid (Bpy-Ala). The incorporation efficiency of BpyRS for Phen-Ala was comparable to that for Bpy-Ala. Because of its high metal-binding ability and characteristic spectral properties, Phen-Ala can be a useful alternative to the existing metal-chelating amino acids for the design and synthesis of artificial metalloproteins.

• Journal of Microbiology and Biotechnology
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• v.26 no.12
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• pp.2019-2029
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• 2016

Zinc finger proteins are among the most extensively applied metalloproteins in the field of biotechnology owing to their unique structural and functional aspects as transcriptional and translational regulators. The classical zinc fingers are the largest family of zinc proteins and they provide critical roles in physiological systems from prokaryotes to eukaryotes. Two cysteine and two histidine residues ($Cys_2His_2$) coordinate to the zinc ion for the structural functions to generate a ${\beta}{\beta}{\alpha}$ fold, and this secondary structure supports specific interactions with their binding partners, including DNA, RNA, lipids, proteins, and small molecules. In this account, the structural similarity and differences of well-known $Cys_2His_2$-type zinc fingers such as zinc interaction factor 268 (ZIF268), transcription factor IIIA (TFIIIA), GAGA, and Ros will be explained. These proteins perform their specific roles in species from archaea to eukaryotes and they show significant structural similarity; however, their aligned amino acids present low sequence homology. These zinc finger proteins have different numbers of domains for their structural roles to maintain biological progress through transcriptional regulations from exogenous stresses. The superimposed structures of these finger domains provide interesting details when these fingers are applied to specific gene binding and editing. The structural information in this study will aid in the selection of unique types of zinc finger applications in vivo and in vitro approaches, because biophysical backgrounds including complex structures and binding affinities aid in the protein design area.

• Radiation Oncology Journal
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• v.33 no.2
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• pp.66-74
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• 2015

Trace elements play crucial role in the maintenance of genome stability in the cells. Many endogenous defense enzymes are containing trace elements such as superoxide dismutase and metalloproteins. These enzymes are contributing in the detoxification of reactive oxidative species (ROS) induced by ionizing radiation in the cells. Zinc, copper, manganese, and selenium are main trace elements that have protective roles against radiation-induced DNA damages. Trace elements in the free salt forms have protective effect against cell toxicity induced by oxidative stress, metal-complex are more active in the attenuation of ROS particularly through superoxide dismutase mimetic activity. Manganese-complexes in protection of normal cell against radiation without any protective effect on cancer cells are more interesting compounds in this topic. The aim of this paper to review the role of trace elements in protection cells against genotoxicity and side effects induced by ionizing radiation.

• Journal of fish pathology
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• v.5 no.2
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• pp.143-152
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• 1992

Metallothionein is an essential and common protein to regulate the intracellular concentration of heavy metals, which exist in most organisms from bacteria to vertebrates. Although the detailed function of metallothianein has not been fully identified until yet, it may be involoved in the cellular protection against the heavy metal toxicity and in the global regulation of several other genes and the expression of metalloproteins. We have cloned the full cDNA clone of metallothionein gene in Channel Catfish by Reverse Transcriptase-Polymerase Chain Reaction(RT-PCR) starting from poly(A)-containing mRNAs. All PCR fragments have been subcloned into EcoRV site of pBluescript SK+ and dT-tailed at Smal site of pUC19, then PCR products are recovered by the double digestion of recombinant plasmids wiht EcoRI and HindIII, which are adjacent to EcoRV site in multicloning sites or by rapid PCR screening. The nucleotide sequence analysis of pMT150(one of the PCR clones) showed high homology with several other piscine metallothionein cDNA genes.

• Korean Journal of Soil Science and Fertilizer
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• v.48 no.5
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• pp.397-403
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• 2015

Heavy metals reduce the photosynthetic efficiency and disrupt metabolic reactions in a concentration-dependent manner. Moreover, by replacing the metal ions in metalloproteins that use essential metal ions, such as Cu, Zn, Mn, and Fe, as co-factors, heavy metals ultimately lead to the formation of reactive oxygen species (ROS). These, in turn, cause destruction of the cell membrane through lipid peroxidation, and eventually cause the plant to necrosis. Given the aforementioned factors, this study was aimed to understand the physiological responses of rice to cadmium (Cd) and arsenic (As) toxicity and the effect of essential metal ions on homeostasis. In order to confirm the level of physiological inhibition caused by heavy metal toxicity, hydroponically grown rice (Oryza sativa L. cv. Dongjin) plants were exposed with $0-50{\mu}M$ cadmium (Cd, $CdCl_2$) and arsenic (As, $NaAsO_2$) at 3-leaf stage, and then investigated malondialdehyde (MDA) contents after 7 days of the treatment. With increasing concentrations of Cd and As, the MDA content in leaf blade and root increased with a consistent trend. At 14 days after treatment with $30{\mu}M$ Cd and As, plant height showed no significant difference between Cd and As, with an identical reduction. However, As caused a greater decline than Cd for shoot fresh weight, dry weight, and water content. The largest amounts of Cd and As were found in the roots and also observed a large amount of transport to the leaf sheath. Interestingly, in terms of Cd transfer to the shoot parts of the plant, it was only transported to upper leaf blades, and we did not detect any Cd in lower leaf blades. However, As was transferred to a greater level in lower leaf blades than in upper leaf blades. In the roots, Cd inhibited Zn absorption, while As inhibited Cu uptake. Furthermore, in the leaf sheath, while Cd and As treatments caused no change in Cu homeostasis, they had an antagonist effect on the absorption of Zn. Finally, in both upper and lower leaf blades, Cd and As toxicity was found to inhibit absorption of both Cu and Zn. Based on these results, it would be considered that heavy metal toxicity causes an increase in lipid peroxidation. This, in turn, leads to damage to the conductive tissue connecting the roots, leaf sheath, and leaf blades, which results in a reduction in water content and causes several physiological alterations. Furthermore, by disrupting homeostasis of the essential metal ions, Cu and Zn, this causes complete heavy metal toxicity.

• Applied Microscopy
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• v.30 no.4
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• pp.347-355
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• 2000

Zinc is one of the most abundant oligoelements in the living cell. It appears tightly bound to some metalloproteins and nucleic acids, loosely bound to some metallothioneins or even as free ion. Small amounts of zinc ions (in the nanomolar range) regulate a plentitude of enzymatic proteins, receptors and transcription factors, thus rolls need accurate homeostasis of zinc ions. Zinc is an essential catalytic or structural element of many proteins, and a signaling messenger that is released by neural activity at many central excitatory synapses. Growing evidences suggest that zinc may also be a key mediator and modulator of the neuronal death associated with transient global ischemia and sustained seizures, as well as perhaps other neurological disease stoles. Some neurons have developed mechanisms to accumulate zinc in specific membrane compartment ('vesicular zinc') which can be evidenced using histochemical techniques. Substances giving a bright colour or emitting fluorescence when in contact with divalent metal ions are currently used to detect them inside cells; their use leads to the so called 'direct' methods. The fixation and precipitation of metal ions as insoluble salt precipitates, their maintenance along the histological process and, finally, their demonstration after autometallographic development are essential steps for other methods, the so called 'indirect methods'. This study is a short report on the autometallograhical approaches for zinc detection in the central nervous system (CNS) by means of a modified selenium method.