• BMB Reports
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• 제37권6호
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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.

• Genomics & Informatics
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• 제15권4호
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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.

• 약학회지
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• 제39권4호
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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.

• 미생물학회지
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• 제32권2호
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• pp.102-108
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• 1994

ABF1(Autonomously replicating sequence Binding Factor 1)은 효모 genome에서 $RTCRYN_5ACG$의 염기 서열을 가지고 있는 promoter, mating-type silencer, ARS에 결합하는 DNA binding 단백질이다. E. coli 에서 ABF1 유전자를 발현하기 위하여, ABF1 유전자를 pMAL-c2 벡터에 cloning하였다.(pMAHW). pMAHW를 E. coli에 형질전환하여, ABF1 융합단백질을 발현시키고, amylose resin affinity chromatography에 의하여 분리하였다. Factor Xa protease를 이용하여 분리된 융합단백질로부터 maltose binding protein을 잘라낸 후에 gel retardation analysis 방법으로 분리된 ABF1이 ARS1에 결합하는 능력을 지니고 있음을 확인하였다. DNA 결합에 관련된 부위를 찾기 위하여, 비전형적인 zinc finger motif가 위치하는 자리에서 pMAHW의 ABF1 유전자에 His-61을 다른 아미노산으로 치환하였다. DNA binding 부위로 추정되는 ABF1 단백질의 중간지역에 Leu-353, Leu-360를 다른 아미노산으로 치환하였다. Site-specific mutagenesis 를 통해 만들어진 mutant를 gel retardation analysis와 complementation test를 통해서 비전형적인 zinc finger motif이외에 다른 DNA binding motif가 있는 것을 알 수 있었다.

• 대한화학회지
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• 제60권2호
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• pp.131-136
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• 2016

CdSe 기반 나노결정의 구리이온에 의한 형광 소광 특성 및 아연이온에 의한 형광 회복 특성을 관찰하였다. 구리이온이 첨가되었을 경우, CdSe 양자점에서는 매우 빠르고 급격한 형광 소광 특성을 보이는 반면에 CdSe 나노라드의 경우에서는 형광이 서서히 소광되는 특성을 보인다. 구리이온으로 형광을 소광시킨 CdSe/CdS(핵/껍질) 양자점에 아연이온이 첨가되면 소광된 양자점의 형광이 회복된다. 용액 내 1 μM의 아연농도에서 양자점의 형광이 50% 증가됨을 확인하였으며, 아연 농도가 증가함에 따라 양자점의 형광세기가 증가되며 이는 Langmuir binding isotherm 모델로 해석할 수 있다. 이러한 연구를 바탕으로 CdSe 기반의 나노결정을 이용한 형광 화학 센서를 구현할 수 있을 것으로 기대한다.

• 생명과학회지
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• 제8권3호
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• pp.235-240
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• 1998

Matrix metalloproteinases(MMP)는 배발생 및 재조직화 등의 정상적인 생체형성과 관절염, 암전이, 치근막염, 골조송증 등의 질병과정에서 collagen이나 proteoglycan과 같은 세포외기질의 구성성분을 분해하는 아연(zinc)효소군이다. 지금까지 다양한 종에서 mmp의 유전자가 클로닝되고 그 기능이 연구되어 왔지만 아직 어류에서는 연구결과가 보고된 바가 없다. 본 연구에서는 한국의 부산연안에 많은 연골어유 투툽상어(Scylliorhinus toraxzame)로부터 RT-PCR(reverse transcriptase dependent polymerase chain reaction)의방법으로 mmp cDNA의 일부를 클로닝하였다. 이것은 염기서열에서 인간, 쥐 및 닭의 membrane type matrix matalloproteinase-3(mt3-mmps)의 염기서열과 74% 동일성을 보이며, 아미노산서열에서는 90%이상의 동일성을 갖고 있다. 또한 MMP에 나타나는 cysteine switch domain, zinc binding domain(HExGH motif), propeptide cleavage site, and RRKR motif등을 가지고 있다. 이러한 결과로부터 본 연구에서 클로닝된 RT-PCR단편은 두툽상어의 mt3-mmp 또는 이와 유사한 유전자의 cDNA이라 믿어진다.

• BMB Reports
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• 제31권1호
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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.

• Preventive Nutrition and Food Science
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• 제9권3호
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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.

• Mycobiology
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• 제36권2호
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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.

• Journal of Genetic Medicine
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• 제16권2호
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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.