• BMB Reports
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• v.48 no.1
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• pp.25-29
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• 2015

Ubiquitination is a post translational modification which mostly links with proteasome dependent protein degradation. This process has been known to play pivotal roles in the number of biological events including apoptosis, cell signaling, transcription and translation. Although the process of ubiquitination has been studied extensively, the mechanism of polyubiquitination by multi protein E3 ubiquitin ligase, SCF complex remains elusive. In the present study, we identified UbcH5a as a novel stimulating factor for poly-ubiquitination catalyzed by $SCF^{hFBH1}$ using biochemical fractionations and MALDI-TOF. Moreover, we showed that recombinant UbcH5a and Cdc34 synergistically stimulate $SCF^{hFBH1}$ catalyzed polyubiquitination in vitro. These data may provide an important cue to understand the mechanism how the SCF complex efficiently polyubiquitinates target substrates.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.1
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• pp.7-12
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• 2000

The expression of the mouse $\alpha-amylase$ gene in the methylotrophic yeast, P pastoris was investigated. The mouse $\alpha-amylase$ gene was inserted into the multi-cloning site of a Pichi a expression vector, pPIC9, yielding a new expression vector pME624. The plasmid pME624 was digested with SalI or BglII, and was introduced into P. pastoris strain GSl15 by the PEG1000 method. Fifty-three transformants were obtained by the transplacement of pME624 digested with SaiII or BglII into the HIS4locus $(38\;of\;Mut^+\;clone)$ or into the AOX1 locus $(15\;of\;Mut^s\;clone)$. Southern blot was carried out in 11 transformants, which showed that the mouse $\alpha-amylase$ gene was integrated into the Pichia chromosome. When the second screening was performed in shaker culture, transformant G2 showed the highest $\alpha-amylase$ activity, 290 units/ml after 3-day culture, among 53 transformants. When this expression level of the mouse $\alpha-amylase$ gene is compared with that in recombinant Saccharomyces cerevisiae harboring a plasmid encoding the same mouse $\alpha-amylase$ gene, the specific enzyme activity is eight fold higher than that of the recombinant S. cerevisiae.

• The Journal of Korean Society of Virology
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• v.30 no.2
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• pp.131-139
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• 2000

Bovine leukemia virus (BLV) is an etiological agent of chronic diseases in cows worldwide. The BLV is one of retroviruses that contain a multi-functional enzyme, reverse transcriptase produced from the pol gene in its genome. We have sequenced some regions in the pol gene of BLV proviruses found in the Southern province of Korea from samples that turned out to be BL V positives by a PCR analysis. On the 5' side of the BLV pol gene (polymerase region), it was found that there were four leucines located at every 7 amino acids. They can form a leucine zipper motif that was not same as the pol gene of Japanese BLV isolate. The sequencing result of the proviral pol gene in Korean-type BLV also revealed some mutations leading to amino acid changes such as $CCT(Pro){\to}CTC(Leu)$, $AAT(Asn){\to}AAA(Lys)$, and non-sensible variations i.e., $TCT(Ser){\to}TCC(Ser)$, $ATT(Ile){\to}ATC(I1e)$ and $ACG(Thr){\to}ACA(Thr)$. On the 3' side of the pol gene (integrase region), some nucleotide sequences were mutated and led to amino acid changes. Among them, a mutation, $GAA(Glu){\to}GAC(Asp)$ occurred in many Korean-type BLV proviruses was very interesting because the amino acid was regarded as one of the most conserved amino acids in the retroviral integrase. It was also notable that the mutation on any leucine residue did not occur, in spite of its frequent appearance.

• International Journal of Oral Biology
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• v.42 no.4
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• pp.191-196
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• 2017

Transglutaminase2 (TGM2) is a multi-functional calcium dependent enzyme that affects angiogenesis, apoptosis, differentiation, attachment, and changes in the extracellular matrix. However, its function in periodontal tissue has not yet been studied. The aim of this study was to investigate the association of the TGM2 expression and the modulation of inflammatory mediators in inflamed periodontal ligament (PDL) cells induced by pro-inflammatory cytokines such as Interleukin-$1{\beta}$ and the Tumor necrosis $factor-{\alpha}$. The expression of TGM2 was increased in the inflamed periodontal tissue and PDL cells. Over-expressed TGM2 in the PDL cells increased expression of MMP1, MMP3, IL-6, CXCL8, and PTGS2. Conversely, inhibition of TGM2 activity using LDN27219, a TGM2 inhibitor, resulted in decreased expression of MMP1, MMP3, IL-6, and CXCL8. The mRNA expression was confirmed by RT-PCR and quantified by qRT-PCR. Protein levels were also confirmed by immunofluoroscence staining. These results suggest that TGM2 plays an important role in the regulation of inflammatory mediators which exacerbate tissue damage in inflamed periodontal tissue.

• Journal of Veterinary Science
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• v.22 no.3
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• pp.29.1-29.6
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• 2021

West Nile virus (WNV), a neurotropic arbovirus, has been detected in mosquitos, birds, wildlife, horses, and humans in Malaysia, but limited information is available on WNV infection in Malaysian pigs. We tested 80 archived swine serum samples for the presence of WNV antibody and West Nile (WN) viral RNA using ID Screen West Nile Competition Multi-species enzyme-linked immunosorbent assay kits and WNV-specific primers in reverse transcription polymerase chain reaction assays, respectively. A WNV seroprevalence of 62.5% (50/80) at 95% confidence interval (51.6%-72.3%) was recorded, with a significantly higher seroprevalence among young pigs (weaner and grower) and pigs from south Malaysia. One sample was positive for Japanese encephalitis virus antibodies; WN viral RNA was not detected in any of the serum samples.

• BioChip Journal
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• v.12 no.4
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• pp.268-279
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• 2018

Flower-shaped organic-inorganic hybrid nanostructures, termed nanoflowers, have received considerable recent attention as they possess greatly enhanced activity, stability, durability, and even selectivity of entrapped organic biomolecules, which are much better than those from the conventional methods. They can be synthesized simply via co-incubation of organic and inorganic components in aqueous buffer at room temperature and yield hierarchical nanostructures with large surface-to-volume ratios, allowing for low-cost production by easy scale-up, as well as the high loading capacity of biomolecules without severe mass transfer limitations. Since a pioneering study reported on hybrid nanoflowers prepared with protein and copper sulfate, many other organic and inorganic components, which endow nanoflowers with diverse functionalities, have been employed. Thanks to these features, they have been applied in a diverse range of areas, including biosensors and biocatalysis. To highlight the progress of research on organic-inorganic hybrid nanoflowers, this review discusses their synthetic methods and mechanisms, structural and biological characteristics, as well as recent representative applications. Current challenges and future directions toward the design and development of multi-functional nanoflowers for their widespread utilization in biotechnology are also discussed.

• Korean Journal of Microbiology
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• v.46 no.2
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• pp.122-126
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• 2010

RAD53 functions as an effector kinase of checkpoint pathways in Saccharomyces cerevisiae, which plays a central role to regulate many downstream cellular processes in response to DNA damage. It also involves in transcriptional activation of various genes including RNR genes which encode the key enzyme required for dNTP synthesis. In this study, we identified CYC8 as a suppressor for the hydroxyurea sensitivity of $rad53{\Delta}$ mutation. $Rad53{\Delta}$ mutant transformed with a multi-copy plasmid containing CYC8 showed increased hydroxyurea resistance. In contrast, TUP1 which forms a complex with CYC8 did not function as a suppressor. In the case of mutations, both $cyc8{\Delta}$ and $tup1{\Delta}$ suppressed hydroxyurea sensitivity of $rad53{\Delta}$. Since CYC8 can propagate as a prion in yeast, overexpression of CYC8 induced misfolding of the normal CYC8 proteins, resulting in dominant cyc8-phenotype. Therefore, it is suggested that CYC8 can act as a multi-copy suppressor due to its prion property. It was observed that the levels of RNR transcription were increased in the yeast strains containing either multi-copies of CYC8 gene or $cyc8{\Delta}$ mutation, suggesting that the increased level of RNR will elevate the intracellular pools of dNTPs, which, in turn, suppress the phenotype of $rad53{\Delta}$ mutation.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.3
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• pp.217-224
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• 2016

Acinetobacter baumannii (A. baumannii) is prevalent in hospital environments and is an important opportunistic pathogen of nosocomial infection. It is known that this pathogen cause herd infection in hospitals, and the mortality rate is remarkably higher for patients infected with this pathogen and already have other underlying diseases. Herein, we investigated the antibiotic resistance rate and the type of resistance genes in 85 isolates of multi-drug resistant A. baumannii from the samples commissioned to laboratory medicine in two university hospitals-in hospital A and hospital B-located in Cheonan and Chungcheong provinces, respectively, in Korea. As a result, $bla_{OXA-23-like}$ and $bla_{OXA-51-like}$ were detected in 82 stains (96.5%). These 82 strains of $bla_{OXA-23-like}$ producing A. baumannii were confirmed with the ISAba1 gene found at the top of the $bla_{OXA-23-like}$ genes by PCR, inducing the resistance against carbapenemase. The armA, AME gene that induces the resistance against aminoglycoside was detected in 34 strains out of 38 strains from Hospital A (89.5%), and in 40 strains out of 47 strains from Hospital B (85.1%), while AMEs were found in 33 strains out of 38 strains from Hospital A (70.2%) and in 44 strains out of 47 strains in Hospital B (93.6%). Therefore, it was found that most multi-drug resistant A. baumannii from the Cheonan area expressed both acethyltransferase and adenyltransferase. This study investigated the multi-drug resistant A. baumannii isolated from Cheonan and Chungcheong provinces in Korea, and it is thought that the results of the study can be utilized as the basic information to cure multi-drug resistant A. baumannii infections and to prevent the spread of drug resistance.

• Korean Journal of Community Nutrition
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• v.12 no.4
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• pp.396-404
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• 2007

Increased oxidative stress contributes to the progression of atherosclerosis. We measured serum antioxidant mineral concentrations, capacities of serum antioxidant enzymes and fasting lipid profile in 97 male patients with coronary artery disease (CAD) and 21 male controls. Nutrient intake was assessed by the semi-quantitative food frequency method. CAD patients were divided into single-vessel disease (SVD, n=66) and multi-vessel disease (MVD, n = 31) groups on the coronary angiography. The ratio of serum LDL- to HDL-cholesterol elevated with an increasing number of diseased vessels compared to the control (control < SVD < MVD, p < 0.05). Patients with SVD and MVD had higher levels of serum lipoprotein (a) than the control (p < 0.05). The mean intake of carbohydrate, protein and cholesterol was higher in MVD patients and the intakes of vitamins C and E were lower in MVD and SVD patients than in the control (p < 0.05). Serum copper (Cu) and zinc (Zn) levels were higher in MVD and SVD patients than in the control (Cu: control $75.8{\pm}5.07$, SVD $99.2{\pm}2.90$, MVD $100.1{\pm}2.32{\mu}g/dL$, p<0.01; Zn: $76.8{\pm}5.36$, $119.0{\pm}5.95$, $129.1{\pm}2.70{\mu}g/dL$, p < 0.01). And the ratio of Zn to Cu was higher in SVD and MVD patients than in the control (control $0.78{\pm}0.06$, SVD $0.88{\pm}0.05$, MVD $0.99{\pm}0.04$, P < 0.05). The activity of glutathione peroxidase (GSH-Px) was lower in MVD than in SVD and the control (control $35.13{\pm}1.34$, SVD $35.30{\pm}1.01$, MVD $31.00{\pm}1.04 U/mg$ protein, p < 0.05). The ratio of the activities of superoxide dismutase (SOD) to GSH-Px was higher in MVD than in control and SVD (p < 0.05). In groups with CAD, serum Cu and Zn concentrations and their ratio were changed compared to the control. GSH-Px activity was decreased and the ratio of SOD to GSH-Px was increased in the patients with MVD. The balances between the activities of SOD and GSH-Px should also be considered a risk factor in CAD patients.

• Korean Journal of Environmental Agriculture
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• v.33 no.4
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• pp.282-289
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• 2014

BACKGROUND: Methane($CH_4$) is considered as the secondmost potent greenhouse gas after carbon dioxide ($CO_2$). Methanogenesis is an enzyme-mediated multi-step process by methanogens. In the penultimate step, methylated Co-M is reduced by methyl Co-M reductase (MCR) to $CH_4$ involving a nickel-containing cofactor F430. The activity of MCR enzyme is dependent on the F430 and therefore, the bioavailability of Ni to methanogens is expected to influence MCR activity and $CH_4$ production in soil. In this study, different doses of EDTA(Ethylene Diamine Tetraacetic Acid) were applied in flooded soils to evaluate their suppression effect on methane production by chelating Ni of methanogenesis cofactor. METHODS AND RESULTS: EDTA was selected as chelating agents and added into wetland and rice paddy soil at the rates of 0, 25, 50, 75, and $100mmol\;kg^{-1}$ before 4-weeks incubation test. During the incubation, cumulative $CH_4$ production patterns were characterized. At the end of the experiment, soil samples were removed from their jars to analyze total soil Ni and water-soluble Ni content and methanogen abundance. Methane production from 100 mmol application decreased by 55 and 78% in both soils compared to that from 0 mmol. With increasing application rate of EDTA in both soils, water-soluble Ni concentration significantly increased, but total soil Ni and methanogen activities showed negative relationship during incubation test. CONCLUSION: The decrease in methane production with EDTA application was caused by chelating Ni of coenzyme F430 and inhibiting methanogenesis by methyl coenzyme M reductase. Consequently, EDTA application decreased uptake of Ni into methanogen, subsequently inhibited methanogen activities and reduced methane production in flooded soils.