• IMMUNE NETWORK
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• v.2 no.4
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• pp.183-188
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• 2002

Gradual attrition of telomere to a critical short length elicits successive cellular response of cellular senescence and crisis. Cancer cells evade this process by maintaining functional telomeres via one of two known mechanisms of telomere maintenance. The first and most frequent mechanism involves reactivation of enzyme activity of telomerase, a ribonucleoprotein complex mainly via transcriptional up-regulation of TERT, a catalytic subunit of telomerase complex. The second mechanism utilizes telomerase-independent way termed ALT (for Alternative Lengthening of Telomere), which possibly involves recombination pathways. Thus master key for cellular immortalization is supposed to possess adequate telomere reserves. Indeed, telomerase can alone induce the immortalization under culture on feeder cell layers without generally known inactivation mechanism of tumor suppressor genes. Including this phenomena, this review will focus on telomerase and telomere-associated proteins, thereby implication of these proteins for cellular immortalization processes.

• BMB Reports
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• v.31 no.5
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• pp.515-518
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• 1998

Adhesive interaction of the platelet glycoprotien IIb-IIIa (GP IIb-IIIa) with a plasma protein, such as fibrinogen, plays an important role in thrombosis and hemostasis. The specific sequence Arg-Gly-Asp (RGD) is critical for the binding of fibrinogen to platelet. To examine and characterize the GP IIb-IIIa antagonist from natural sources, we have developed a simple enzyme-linked immunosorbant assay (ELISA) system. The GP IIb-IIIa complex was purified to homogeneity from platelet Iysates by the combination of two affinity chromatographic methods using the synthetic RGD peptide (GRGDSPK)-immobilized Sepharose and wheat germ lectin-Sepharose. The synthetic peptide GRGDSP inhibits GP IIb-IIIa binding to immobilized fibrinogen with an $IC_{50}$ of $1.5\;{\mu}M$. Venoms of three different snake species and a Korean scolopendra extract have strong antagonistic activities for the binding of human fibrinogen to the platelet GP IIb-IIIa complex. The $IC_{50}$ values of the snake venom s and scolopendra were in the range of $5.5\;{\mu}g$ to $60\;{\mu}g$. These results provide meaningful information for developing antiplatelet agents.

• Journal of Electrochemical Science and Technology
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• v.9 no.3
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• pp.169-175
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• 2018

Cu-Salen complex was prepared and attached into chitosan (Cs) polymer backbone. Nanocomposite of the synthesized polymer was prepared with functionalized carbon nano-particles (Cs-Cu-sal/C) to modify the electrode surface. The surface morphology of (Cs-Cu-sal/C) nanocomposite film showed a homogeneous distribution of carbon nanoparticles within the polymeric matrix. The cyclic voltammogram of the modified electrode exhibited a redox behavior at -0.1 V vs. Ag/AgCl (3 M KCl) in 0.1 M PB (pH 7) and showed an excellent hydrogen peroxide reduction activity. The Cs-Cu-sal/C electrode displays a linear response from $5{\times}10^{-6}$ to $5{\times}10^{-4}M$, with a correlation coefficient of 0.993 and detection limit of $0.9{\mu}M$ (at S/N = 3). The sensitivity of the electrode was found to be $0.356{\mu}A\;{\mu}M^{-1}\;cm^{-2}$.

• Journal of the Korean Society of Food Science and Nutrition
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• v.22 no.6
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• pp.833-838
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• 1993

A strain of Penicillium sp. extracellularly produced an inhibitory substance for lipoxygenase. These purification procedures were followed : ethanol treatment, chromatographies on Dowex 50W, Sephadex G-25, silica gel column and HPLC. The inhibitor was stable in pH range from 3.0 to 5.0 at $25^{\circ}C$, and a treatment at 10$0^{\circ}C$ for 2 hours didn't diminish its original activity. The purified inhibitor was charred at temperature near 22$0^{\circ}C$~23$0^{\circ}C$ and decomposed. Molecular weight of the inhibitor was estimated to be approximately 270 by Sephadex G-25 column chromatography. The inhibitor rapidly formed EI complex with lipoxygenase and inhibited enzyme activity.

• Archives of Pharmacal Research
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• v.28 no.7
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• pp.761-764
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• 2005

Indigin, alkylated xanthene (1) and indigoferic acid (2) have been isolated from the chloroform soluble fraction of Indigofera oblongifolia, along with $\beta$-sitosterol (3) and 3-hydroxybenzoic acid (4), which are reported for the first time from this species. Their structures were determined through spectroscopic techniques. Both the new compounds 1 and 2 showed significant activity against enzyme lipoxygenase, while 2 further showed moderate inhibition against BChE.

• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.656-662
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• 2000

A high level of Staphylococcus haemolyticus L62 lipase was expressed in an Escherichia coli transformant. The expressed lipase activity in the cell-free extract was 70,800 U/l, which corresponded to 30% of the total cellular protein. Pre-mixing of the l62 lipase with some nonionic detergents enhanced its hydrolytic activity towards olive oil: Tween detergents activated the L62 lipase by 3 fold. Gel filtration chromatography of the Tween-80-L62 lipase mixture demonstrated a polymerized complex (∼180 kDa) formed exclusively between Tween-80 and the L62 lipase. The lipase enzyme in the complex showed a higher specific activity towards most triacylglycerols than the intact L62 lipase. The activity enhancement towards each substrate was quite different depending on the acyl chain length; the activity towards tributyrin, trilinolein, and trilinolenin was much more enhanced than the towards the medium and the long-chain saturated triglycerides.

• Korean Journal of Food Science and Technology
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• v.36 no.5
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• pp.765-772
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• 2004

Cooking condition of Tarakjuk (milk-rice porridge) was established based on gelatinization temperature using differential scanning calorimetry (DSC) of roasted Ilpum rice flour, which has highest enzyme-resistant starch (RS) content. Effect of cooking temperature and time on DSC characteristics, crystallity with X ray diffractogram, RS content, in vitro starch digestibility (IVSD), amino acid composition, and in vitro protein digestibility (IVPD) of Tarakjuk were determined. Tarakjuk was cooked at 50, 56.5, 64, and $69^{\circ}C$ for various durations. Rice flour ingredient used was Ilpum, previously roasted at $185^{\circ}C$ for 25 min. Tarakjuk cooked at 50 and $56.5^{\circ}C$ showed two thermal transitions between $63.7-125.2^{\circ}C$ as determined by DSC, corresponding to endotherms of starch gelatinization $(63.7-73.8^{\circ}C)$ and melting of amylose-lipid complex (AM-lipid complex, $97.7-125.2^{\circ}C$), whereas that cooked at 64 and $69^{\circ}C$ showed only AM-lipid complex melting transition between $96.9-127.6^{\circ}C$. As cooking temperature increased, RS content of Tarakjuk decreased, whereas IVSD increased. Total amino acid content was between 11,558-15,601mg/100g, depending on cooking condition used. Compared with conventionally made control, contents of essential amino acids, such as lysine and tryptophane, were higher at 50 and $56.5^{\circ}C$, and IVPD showed higher increase. Results reveal degree of gelatinization in Tarakjuk with high RS content as well as low IVSD and high IVPD, which are important from physiological and nutritional point of view, can be produced by controlling cooking condition.

• The Korean Journal of Zoology
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• v.39 no.1
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• pp.36-46
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• 1996

An endogenous phenoloxidase (EPO) from earthworm, Lumbricus rubellus, has been purified and characterized. The purified EPO using ammonium sulfate fractionation, Blue-2, Phenyl-, and Q-sepharose chromatography steps was revealed in SDS-PAGE as a single protein banri with Mr. of 59 kl)a. A native strudure of the enzyme was examined with an in situ staining of a nondenatudng-PAGE using DL-dopa as a substrate. The result showed that a single band due to the EPO activity was located siighdy above a standard polypeptide with Mr. of 210 kl)a. These fads indicate that the EPO is an oligomeric enzyme. The presence of a monophenolase activity of the purified EPO, which hydroxylates tyrosine to dopa, was confirmed by observing dopachrome accumulation at 475 nm at PH 8.0 with a typical lag phase during 60 mm. of meausrement. A series of inhibition study has been performed for the enzyme with several divalent cation chelators such as phenyithiourea (Flu), 1, lO-phenanthroline, EDTA, and EGTA. Among them, only V'flj inhibited the enzyme with 1C0.5 of 65 MM, which indicated that copper was critical for the catalysis of EPO. The enzyme was maximally active at 35'C and pH 8.0 when L-dopa to dopachrome conversion was spectrophotometricaily monitored at 475 nm. The apparent Km values of P0 for L-opa were obtained as 1.86 mM and 13.8 mM at pH 6.5 and 8.0, respectively. The catalytic efficiencies at both pH were almost identical [(kat/Km)pH8.0/(kcat/Km)pH6.5 = O.92] while the Vmax at p11 8.0 was 6.6-fold higher than that at pH 6.5. This fact may indicate that pH affeds the catalysis at substrate and/or enzyme-substrate complex level rather than the enzyme itself. Taken together, the EPO was an oligomeric enzyme which did not require proteolysis for its activation. These results also indicated that the enzyme can exist, at least, in part as a latent form In vivo, which might be distinct from the prophenoloxidase activating system. Therefore, it is pertinent to consider that there must be certain regulatory molecules or phenomena in L. rubellus which make the 1,0 in a latent form in vivo before the foreign invasions.

• Bulletin of the Korean Chemical Society
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• v.34 no.9
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• pp.2671-2674
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• 2013

To elucidate the role of the evolutionarily conserved Tyr8 residue in rice Phi-class GSTF3, this amino acid was replaced with alanine and phenylalanine by site-directed mutagenesis, respectively. The replacement of Tyr8 with Ala significantly affected the catalytic activity and the kinetic parameters, whereas the substitutions of Tyr8 with Phe had almost no effect. The Y8A mutant resulted in approximately 90-100% decrease of the specific activity. Moreover, the Y8A mutant resulted approximately in 2-fold increase of $K_m$, approximately 60-80% decrease of $k_{cat}$, and approximately 6.5-fold decrease in $k_{cat}/K_m$. From the pH/log $k_{cat}/K_m$ plot, $pK_a$ values of the GSH in the wild-type enzyme-GSH complex, Y8A-GSH complex and Y8F-GSH complex were estimated to be approximately 6.8, 8.5 and 6.9, respectively. From these results, we suggest that the evolutionarily conserved Tyr8 residue in OsGSTF3 seems to influence the structural stability of the active site of OsGSTF3 rather than directly its catalytic activity.

• Molecules and Cells
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• v.39 no.9
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• pp.669-679
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• 2016

N-acetyl-D-glucosamine kinase (GlcNAc kinase or NAGK) primarily catalyzes phosphoryl transfer to GlcNAc during amino sugar metabolism. Recently, it was shown NAGK interacts with dynein light chain roadblock type 1 (DYNLRB1) and upregulates axo-dendritic growth, which is an enzyme activity-independent, non-canonical structural role. The authors examined the distributions of NAGK and NAGK-dynein complexes during the cell cycle in HEK293T cells. NAGK was expressed throughout different stages of cell division and immunocytochemistry (ICC) showed NAGK was localized at nuclear envelope, spindle microtubules (MTs), and kinetochores (KTs). A proximity ligation assay (PLA) for NAGK and DYNLRB1 revealed NAGK-dynein complex on nuclear envelopes in prophase cells and on chromosomes in metaphase cells. NAGK-DYNLRB1 PLA followed by Lis1/NudE1 immunostaining showed NAGK-dynein complexes were colocalized with Lis1 and NudE1 signals, and PLA for NAGK-Lis1 showed similar signal patterns, suggesting a functional link between NAGK and dynein-Lis1 complex. Subsequently, NAGK-dynein complexes were found in KTs and on nuclear membranes where KTs were marked with CENP-B ICC and nuclear membrane with lamin ICC. Furthermore, knockdown of NAGK by small hairpin (sh) RNA was found to delay cell division. These results indicate that the NAGK-dynein interaction with the involvements of Lis1 and NudE1 plays an important role in prophase nuclear envelope breakdown (NEB) and metaphase MT-KT attachment during eukaryotic cell division.