• Title/Summary/Keyword: zinc binding

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Identification of the DNA Binding Element of the Human ZNF333 Protein

  • Jing, Zhe;Liu, Yaping;Dong, Min;Hu, Shaoyi;Huang, Shangzhi
    • BMB Reports
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    • v.37 no.6
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    • pp.663-670
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    • 2004
  • ZNF 333 is a new and sole gene containing two KRAB domains which has been identified currently. It is a member of subfamilies of zinc finger gene complex which had been localized on chromosome 19p13.1. The ZNF333 gene mainly encodes a 75.5 kDa protein which contains 10 zinc finger domains. Using the methods of random oligonucleotide selection assay, electromobility gel shift assay and luciferase activity assay, we found that ZNF333 recognized the specific DNA core binding sequence ATAAT. Moreover, these data indicated that the KRAB domain of ZNF333 really has the ability of transcriptional repression.

Prediction of Metal Ion Binding Sites in Proteins from Amino Acid Sequences by Using Simplified Amino Acid Alphabets and Random Forest Model

  • Kumar, Suresh
    • Genomics & Informatics
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    • v.15 no.4
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    • pp.162-169
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    • 2017
  • Metal binding proteins or metallo-proteins are important for the stability of the protein and also serve as co-factors in various functions like controlling metabolism, regulating signal transport, and metal homeostasis. In structural genomics, prediction of metal binding proteins help in the selection of suitable growth medium for overexpression's studies and also help in obtaining the functional protein. Computational prediction using machine learning approach has been widely used in various fields of bioinformatics based on the fact all the information contains in amino acid sequence. In this study, random forest machine learning prediction systems were deployed with simplified amino acid for prediction of individual major metal ion binding sites like copper, calcium, cobalt, iron, magnesium, manganese, nickel, and zinc.

Biochemical Characteristics of Cd-binding High Molecular Weight Proteins (카드뮴 결합 고분자량 단백질의 생화학적 특성)

  • 천기정;김봉희
    • YAKHAK HOEJI
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    • v.39 no.4
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    • pp.380-384
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    • 1995
  • The isoelelectric points of Cd-BP(l) and Cd-BP(II), cadmium-binding proteins, were 6.01 and 5.35, respectively. Both of them contained zinc. As for the amino acid composition, Cd-BP(I) contained a lot of glycine and lysine but none of such aromatic amino acids as tyrosine and phenylalanine.. On the other hand, Cd-BP(II) contained leucine, histidine, asparti cacid and alanine but no aromatic amino acids.

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Structure-Function Analysis of DNA Binding Domain of the Yeast ABF1 Protein (효모 ABF1 단백질의 DNA Binding 부위에 대한 구조 기능 연구)

  • Cho, Gi-Nam;Lee, Sang-Kyung;Kim, Hong-Tae;Kim, Ji-Young;Rho, Hyune-Mo;Jung, Gu-Hung
    • Korean Journal of Microbiology
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    • v.32 no.2
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    • pp.102-108
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    • 1994
  • Autonomously replicating sequence Binding Factor 1(ABF1) is a DNA-binding protein that specifically recognizes the $RTCRYN_5ACG$ at many sites in the yeast genome including the promoter element, mating-type silencer and ARS. To express the intact full-length ABF1 gene in E. coli, the ABF1 gene has been cloned into pMAL-c2 and His-61, Leu-353 and Leu-360 were substituted with other amino acid. ABF1 fusion proteins of wild type ABF1 and H61A, L353R and L360R nutants were purified by amylose resin affinity chromatography. Fusion protein of MBP and ABF1 was digested by Factor Xa and Characterized by gel retardation assay and complementation test. As aresult, we suggested that other DNA binding motif except atypical inc-finger motif is in the middle region of ABF1.

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Photoluminescence Quenching and Recovery of the CdSe Nanocrystals by Metal Ions (금속이온에 의한 CdSe 나노결정의 형광 소광 및 회복 특성)

  • Bang, Jiwon;Kim, Bomi;Koo, Eunhae;Kim, Sungjee
    • Journal of the Korean Chemical Society
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    • v.60 no.2
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    • pp.131-136
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    • 2016
  • Copper ion induced photoluminescence (PL) quenching dynamics and recovery of the PL by zinc ions were investigated for CdSe based nanocrystals. When copper ions were added, CdSe quantum dots showed fast and dramatically PL quenching whereas PL of CdSe nanorod gradually decreased. In the presence of zinc ions, the PL of CdSe/CdS (core/shell) nanocrystals that have quenched by copper ions was efficiently recovered. It showed that the PL intensity of nanocrystals increased by 50% in a solution containing 1 μM zinc ions. The PL intensity was increasing with increasing zinc ions, and could be described by Langmuir binding isotherm model. We showcase that the CdSe based nanocrystals can be used as fluorescence turn-on sensor.

Cloning of a matrix metalloproteinase cDNA from Scylliorhinus torazame (두툽상어 matrix metalloproteinase 유전자 cDNA의 클로닝)

  • Kim, Jon Won;Cho, Won Jin;Chun, Kwang Ho;Kim, Kyu-Won;Kim, Yung-Jin;Lee, Sang-Jun;Shin, Hae-Ja;Lim, Woon Ki
    • Journal of Life Science
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    • v.8 no.3
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    • pp.235-240
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    • 1998
  • Matrix metalloproteinases(MMPs) are a group of zinc enzymes responsible for degradation of the matrix components such as collagen and proteoglycans in normal embryogenesis and remodeling and in many disease processes such as arthritis, cancer, periodontitis, and osteprocess. Genetically distince MMPs have been characterized and their genes have been cloned thus far from a variaty of species but not from fishes. In this stydy, a mmp cDNA was cloned by using RT-PCR(reverse transcriptase dependent polymerase chain reaction) from Scylliorhinus toraxzame(shark), agroup of cartilaginous fish, abundant in the coast of Pusan, Korea. It has 74% base homologue with membrane type matrix matalloproteinase-3 genes(mt3-mmps) from human, rat and chick, and also shows more than 90% residue homologue with them. In addition, it has cysteine switch domain, zinc binding domain(HExGH motif), propeptide cleavage site, and RRKR motif, which are present in MMPs. This result indicates that cDNA fragment cloned here may be mt3-mmp or its analogous gejne cDNA fragment of Scylliorhinus torzame.

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Unusual Allosteric Property of L-alanine Dehydrogenase from Bacillus subtilis

  • Kim, Soo-Ja;Lee, Woo-Yiel;Kim, Kwang-Hyun
    • BMB Reports
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    • v.31 no.1
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    • pp.25-30
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    • 1998
  • Kinetic studies of L-Alanine dehydrogenase from Bacillus subtilis-catalyzed reactions in the presence of $Zn^{2+}$ were carried out. The substrate (L-alanine) saturation curve is hyperbolic in the absence of the metal ion but it becomes sigmoidal when $Zn^{2+}$ is added to the reaction mixture indicating the positive cooperative binding of the substrate in the presence of zinc ion. The cooperativity of substrate binding depends on the xinc ion concentration: the Hill coefficients ($n_H$) varied from 1.0 to 1.95 when the zinc ion concentration varied from 0 to $60\;{\mu}m$. The inhibition of AlaDH by $Zn^{2+}$ is reversible and noncompetitive with respect to $NAD^+$ ($K_i\;=\;5.28{\times}10^{-5}\;M$). $Zn^{2+}$ itself binds to AlaDH with positive cooperativity and the cooperativity is independent of substrate concentration. The Hill coefficients of substrate biding in the presence of $Zn^{2+}$ are not affected by the enzyme concentration indicating that $Zn^{2+}$ binding does not change the polymerization-depolymerization equilibria of the enzyme. Among other metal ions, $Zn^{2+}$ appears to be a specific reversible inhibitor inducing conformational change through the intersubunit interaction. These results indicate that $Zn^{2+}$ is an allosteric competitive inhibitor and substrate being a non-cooperative per se, excludes the $Zn^{2+}$ from its binding site and thus exhibits positive cooperativity. The allosteric mechanism of AlaDh from Bacillus subtilis is consistent with both MWC and Koshland's allosteric model.

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Zinc Status Assessment by Analysis of Mononuclear Cell Metallothionein mRNA Using Competitive-Reverse Transcriptase-Polymerase Chain Reaction

  • Lee, Soo-Lim;Yoon, Jin-Sook;Kwon, Chong-Suk;Beattie, John H.;Kwun, In-Sook
    • Preventive Nutrition and Food Science
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    • v.9 no.3
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    • pp.276-282
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    • 2004
  • Marginal Zn deficiency is prevalent through the world and yet human zinc status has not been properly assessed due to the lack of a reliable diagnostic indicator. One potential possibility for zinc status assessment using Zn-binding protein, metallothionein (MT)-mRNA, has been proposed. The purpose of the present study was aimed to show whether measurement of mononuclear cell (MNC) MT mRNA, using a competitive-reverse transcriptase-polymerase chain reaction (competitive-RT-PCR) assay, could indicate zinc status in human subjects. In this study, MNC MT-mRNA expression was measured using a competitive-RT-PCR to compare before and after 14 days of zinc supplementation (50 mg Zn/das zinc gluconate). RT-PCR oligonucleotide primers which were designed to amplify both a 278 bp segment of the human MT-2A cDNA and a 198 bp mutant competitor cDNA template from MNCs, were prepared. MT-2A mRNA was normalized by reference to the housekeeping gene, $\beta$-actin, mRNA for which was also measured by competitive-RT-PCR. There was considerable inter-individual variation in MT-mRNA concentration and yet, the mean MT-2A mRNA level increased 4.7-fold after Zn supplementation, as compared to before Zn supplementation. This MT-2A mRNA level was shown as the same pattern and, even more sensitive assay, compared to the conventional plasma and red blood cells (RBCs) Zn assessment in which plasma and RBCs zinc levels increased 2.3- and 1.2-fold, respectively (p<0.05). We suggest that MT competitive-RT-PCR can be a useful assessment tool for evaluating human zinc status.

Antibacterial and Antifungal Studies on Some Schiff Base Complexes of Zinc(II)

  • Joseyphus, R. Selwin;Nair, M. Sivasankaran
    • Mycobiology
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    • v.36 no.2
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    • pp.93-98
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    • 2008
  • Two Schiff base ligands $L_1\;and\;L_2$ were obtained by the condensation of glycylglycine respectively with imidazole-2-carboxaldehyde and indole-3-carboxaldehyde and their complexes with Zn(II) were prepared and characterized by microanalytical, conductivity measurement, IR, UV-Vis., XRD and SEM. The molar conductance measurement indicates that the Zn(II) complexes are 1:1 electrolytes. The IR data demonstrate the tetradentate binding of $L_1$ and tridentate binding of $L_2$. The XRD data show that Zn(II) complexes with $L_1\;and\;L_2$ have the crystallite sizes of 53 and 61 nm respectively. The surface morphology of the complexes was studied using SEM. The in vitro biological screening effects of the investigated compounds were tested against the bacterial species Staphylococcus aureus, Escherichia coli, Klebsiella pneumaniae, Proteus vulgaris and Pseudomonas aeruginosa and fungal species Aspergillus niger, Rhizopus stolonifer, Aspergillus flavus, Rhizoctonia bataicola and Candida albicans by the disc diffusion method. A comparative study of inhibition values of the Schiff base ligands and their complexes indicates that the complexes exhibit higher antimicrobial activity than the free ligands. Zinc ions are proven to be essential for the growth-inhibitor effect. The extent of inhibition appeared to be strongly dependent on the initial cell density and on the growth medium.

Periventricular nodular heterotopia in a child with a mild Mowat-Wilson phenotype caused by a novel missense mutation of ZEB2

  • Kim, Young Ok;Lee, Yun Young;Kim, Myeong-Kyu;Woo, Young Jong
    • Journal of Genetic Medicine
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    • v.16 no.2
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    • pp.71-75
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    • 2019
  • Periventricular nodular heterotopia (PNH) is a malformation of cortical development in which normal neurons inappropriately cluster in periventricular areas. Patients with Mowat-Wilson syndrome (MWS) typically present with facial gestalt, complex neurologic problems (e.g., severe developmental delay with marked speech impairment and epilepsy), and multiple anomalies (e.g., Hirschsprung disease, urogenital anomalies, congenital heart defects, eye anomalies, and agenesis of the corpus callosum [CC]). MWS is mostly caused by haploinsufficiency of the gene encoding zinc-finger E-box-binding homeobox 2 (ZEB2) due to premature stops or large deletions. We present a case report of a 9-year-old girl with PNH, drug-responsive epilepsy, severe intellectual disability, and facial dysmorphisms only in whom we performed whole-exome sequencing and found a de novo heterozygous missense mutation (c.3134A>C; p.His1045Pro) of ZEB2 (NM_014795.3; NP_055610.1). This mild case of MWS caused by a rare novel missense mutation of ZEB2 represents the first report of MWS with isolated PNH.