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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Korean Society for Biochemistry and Molecular Biology
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Volume & Issues
Volume 36, Issue 6 - Nov 2003
Volume 36, Issue 5 - Sep 2003
Volume 36, Issue 4 - Jul 2003
Volume 36, Issue 3 - May 2003
Volume 36, Issue 2 - Mar 2003
Volume 36, Issue 1 - Jan 2003
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Theoretical Investigation of the Triphosphate Forms of Azidothymidine and Thymidine
Arissawa, Marcia ; Felcman, Judith ; Herrera, Juan Omar Machucca ;
BMB Reports , volume 36, issue 3, 2003, Pages 243~250
DOI : 10.5483/BMBRep.2003.36.3.243
In this paper we investigate (using AM1 semi-empirical as well as HF methods at the STO-3G, 3-21G, 6-31G, 6-
level) the conformations, geometrical parameters, Mulliken charges, and solvation effects of the triphosphate form of AZT (AZTTP), as well as the thymidine nucleotide (dTTP) structure. Our calculated geometrical parameters and Mulliken charges, with and without solvation effects, are correlated with recent experimental results.
Agmatine Reduces Hydrogen Peroxide in Mesangial Cells under High Glucose Conditions
Lee, Geun-Taek ; Ha, Hun-Joo ; Lee, Hyun-Chul ; Cho, Young-Dong ;
BMB Reports , volume 36, issue 3, 2003, Pages 251~257
DOI : 10.5483/BMBRep.2003.36.3.251
Agmatine, an amine and organic cation, reduced
that was generated by hyperglycemia, and transcription factors such as NF-
and AP-1 activity in the mesangial cells that were exposed to high glucose. However, spermine which shares a strong nucleophilic structure with agmatine decreased the
levels and AP-1, but not the NF-
activity. Possible roles for agmatine and spermine in decreasing fibronectin are discussed, and the signaling pathway for agmatine-reduced fibronectin accumulation is presented.
Suppression of Fatty Acid Synthase by Dietary Polyunsaturated Fatty Acids is Mediated by Fat itself, not by Peroxidative Mechanism
Kim, Hye-Kyeong ; Choi, Sung-Won ; Lee, Hae-Jeung ; Lee, Joo-Hee ; Choi, Hay-Mie ;
BMB Reports , volume 36, issue 3, 2003, Pages 258~264
DOI : 10.5483/BMBRep.2003.36.3.258
This study examined the effect of dietary polyunsaturated fatty acids (PUFA) that were supplemented with vitamin E on lipid peroxidation, glutathione-dependent detoxifying enzyme system activity, and lipogenic fatty acid synthase (FAS) expression in rat liver. Male Sprague-Dawley rats were fed semipurified diets containing either 1% (w/w) corn oil or 10% each of beef tallow, corn oil, perilla oil, and fish oil for 4 wk. Alpha-tocopherol was supplemented in perilla oil (0.015%) and fish oil (0.019%). Hepatic thiobarbituric acid reactive substances, an estimate of lipid peroxidation, were not significantly different among the dietary groups. The glutathione peroxidase, glutathione reductase, and glutathione S-transferase activities were all elevated by the polyunsaturated fats, especially fish oil. The activity of FAS was reduced in the polyunsaturated fat-fed groups in the order of fish oil, perilla oil, and corn oil. The mRNA contents decreased in rats that were fed the 10% fat diets, particularly polyunsaturated fats, compared with the rats that were fed the 1% corn oil diet. Similarly, the inhibitory effect was the greatest in fish oil. These results suggest that lipid peroxidation can be minimized by vitamin E; PUFA in itself has a suppressive effect on lipogenic enzyme.
Retinoic Acid-Induced Golgi Apparatus Disruption in F2000 Fibroblasts: A Model for Enhanced Intracellular Retrograde Transport
Tzankov, Alexandar ;
BMB Reports , volume 36, issue 3, 2003, Pages 265~268
DOI : 10.5483/BMBRep.2003.36.3.265
Retinoic acid (RA) can transform the Golgi apparatus (GA) into a diffuse vacuolar aggregate and increase the toxicity of some immunotoxins that enter into cells by receptor-mediated endocytosis. An ultramorphological study of the RA-induced GA disruption was performed on F2000 fibroblasts. Cultures were treated with 0.11 to
RA for 7 - 180 min. The endocytosis of Limax flavus agglutinin-peroxidase conjugate (LFA), and the interactions between a phorbol ester (PMA) and RA concerning GA disruption, were examined. Exposure to
RA for 20 min transformed the GA into vacuolar aggregate. These vacuoles were not involved in endocytosis since they remained unstained after endocytosis of LFA. However, the lysosomes were involved in endocytosis, as they were strongly stained. Therefore, a RA-induced shift towards lysosomal routing of the entered LFA was presumed. Exposure to PMA made cells resistant to the Golgi-disturbing effects of RA, indicating that protein kinase C plays an important role in this process.
Styrylpyrone Derivative Induces Apoptosis through the Up-Regulation of Bax in the Human Breast Cancer Cell Line MCF-7
Chien, Alvin Lee Teck ; Pihie, Azimahtol Hawariah Lope ;
BMB Reports , volume 36, issue 3, 2003, Pages 269~274
DOI : 10.5483/BMBRep.2003.36.3.269
In the fight against cancer, novel chemotherapeutic agents are constantly being sought to complement existing drugs. Various studies have presented evidence that the apoptosis that is induced by these anticancer agents is implicated in tumor regression, and Bcl-2 family genes play a part in apoptosis following treatment with various stimuli. Here, we present data that a styrylpyrone derivative (SPD) that is extracted from the plant Goniothalamus sp. showed cytotoxic effects on the human breast cancer cell line MCF-7. SPD significantly increased apoptosis in MCF-7 cells, as visualized by phase contrast microscopy and evaluated by the Tdt-mediated dUTP nick end-labeling assay and nuclear morphology. Western blotting and immunostaining revealed up-regulation of the proapoptotic Bax protein expression. SPD, however, did not affect the expression of the anti-apoptotic protein, Bcl-2. These results, therefore, suggest SPD as a potent cytotoxic agent on MCF-7 cells by inducing apoptosis through the modulation of Bax levels.
Conditional Replication of a Recombinant Adenovirus Studied Using Neomycin as a Selective Marker
Xue, Feng ; Qi, Yi-Peng ; Joshua, Mallam Nock ; Lan, Ping ; Dong, Chang-Yuan ;
BMB Reports , volume 36, issue 3, 2003, Pages 275~281
DOI : 10.5483/BMBRep.2003.36.3.275
An E1B-defective adenovirus, named r2/Ad carrying the neo expression cassette, was constructed by homologous recombination. The construction, selection (using neomycin as a selective marker), and propagation of the recombinant virus was performed in human embryonic kidney 293 cells (HEK 293). An in vitro study demonstrated that this recombinant virus has the ability to replicate in and lyse some p53-deficient human tumor cells such as human glioma tumor cells (U251) and human bladder cells (EJ), but not in some cells with functional p53, such as human adenocarcinoma cells (A549) and human fibroblast cells (MRC-5). Also, based on the cytopathic effect (CPE), it was demonstrated, under identical conditions, that the U251 cells were more sensitive to r2/Ad replication than the EJ cells. In this paper, we report that r2/Ad could be very useful in studying the in vitro selective replication of E1B-defective adenovirus and has great potential in cancer gene therapy.
Production of Superoxide Dismutase by Deinococcus radiophilus
Yun, Young-Sun ; Lee, Young-Nam ;
BMB Reports , volume 36, issue 3, 2003, Pages 282~287
DOI : 10.5483/BMBRep.2003.36.3.282
The production of superoxide dismutase (SOD) varied in Deinococcus radiophilus, the UV resistant bacterium, depending upon different phases of growth, UV irradiation, and superoxide treatment. A gradual increase in total SOD activity occurred up to the stationary phases. The electrophoretic resolution of the SOD in cell extracts of D. radiophilus at each growth phase revealed the occurrence of MnSOD throughout the growth phases. The SOD profiles of D. radiophilus at the exponential phase received oxidative stress by the potassium superoxide treatment or UV irradiation also revealed the occurrence of a single SOD. However, these treatments caused an increase in SOD activity. The data strongly suggest that D. radiophilus has only one species of SOD as a constitutive enzyme, which seems to be a membrane-associated protein.
The EphA8 Receptor Phosphorylates and Activates Low Molecular Weight Phosphotyrosine Protein Phosphatase in Vitro
Park, Soo-Chul ;
BMB Reports , volume 36, issue 3, 2003, Pages 288~293
DOI : 10.5483/BMBRep.2003.36.3.288
Low molecular weight phosphotyrosine protein phosphatase (LMW-PTP) has been implicated in modulating the EphB1-mediated signaling pathway. In this study, we demonstrated that the EphA8 receptor phosphorylates LMW-PTP in vitro. In addition, we discovered that mixing these two proteins leads to EphA8 dephosphorylation in the absence of phosphatase inhibitors. Finally, we demonstrated that LMW-PTP, modified by the EphA8 autokinase activity, possesses enhanced catalytic activity in vitro. These results suggest that LMW-PTP may also participate in a feedback-control mechanism of the EphA8 receptor autokinase activity in vivo.
Ex vivo Cytotoxicity of the Bacillus thuringiensis Cry4B δ-Endotoxin to Isolated Midguts of Aedes aegypti Larvae
Barusrux, Sahawat ; Sramala, Issara ; Katzenmeier, Gerd ; Bunyaratvej, Ahnond ; Panyim, Sakol ; Angsuthanasombat, Chanan ;
BMB Reports , volume 36, issue 3, 2003, Pages 294~298
DOI : 10.5483/BMBRep.2003.36.3.294
The pathological effect of the Bacillus thuringiensis Cry
-endotoxins on susceptible insect larvae had extensive damage on the midgut epithelial cells. In this study, an ex vivo assay was devised for assessing the insecticidal potency of the cloned Cry4B mosquito-larvicidal protein that is expressed in Escherichia coli. Determination of toxicity was carried out by using a cell viability assay on the midguts that were dissected from 5-day old Aedes aegypti mosquito larvae. After incubation with the toxin proteins, the number of viable epithelial cells was determined photometrically by monitoring the quantity of the bioreduced formazan product at 490 nm. The results showed that the 65-kDa trypsin-activated Cry4B toxin exhibited toxic potency ca. 3.5 times higher than the 130-kDa Cry4B protoxin. However, the trypsin-treated products of the non-bioactive Cry4B mutant (R158A) and the lepidopteran-specific Cry1Aa toxin displayed relatively no ex vivo activity on the mosquito-larval midguts. The ex vivo cytotoxicity studies presented here confirms data that was obtained in bioassays.
Identification of Novel Target Proteins of Cyclic GMP Signaling Pathways Using Chemical Proteomics
Kim, Eui-Kyung ; Park, Ji-Man ;
BMB Reports , volume 36, issue 3, 2003, Pages 299~304
DOI : 10.5483/BMBRep.2003.36.3.299
For deciphering the cyclic guanosine monophosphate (cGMP) signaling pathway, we employed chemical proteomics to identify the novel target molecules of cGMP. We used cGMP that was immobilized onto agarose beads with linkers directed at three different positions of cGMP. We performed a pull-down assay using the beads as baits on tissue lysates and identified 9 proteins by MALDI-TOF (Matrix-Assisted Laser Desorption/Ionization Time-of-Flight) mass spectrometry. Some of the identified proteins were previously known cGMP targets, including cGMP-dependent protein kinase and cGMP-stimulated phosphodiesterase. Surprisingly, some of the co-precipitated proteins were never formerly reported to associate with the cGMP signaling pathway. The competition binding assays showed that the interactions are not by nonspecific binding to either the linker or bead itself, but by specific binding to cGMP. Furthermore, we observed that the interactions are highly specific to cGMP against other nucleotides, such as cyclic adenosine monophosphate (cAMP) and 5'-GMP, which are structurally similar to cGMP. As one of the identified targets, MAPK1 was confirmed by immunoblotting with an anti-MAPK1 antibody. For further proof, we observed that the membrane-permeable cGMP (8-bromo cyclic GMP) stimulated mitogen-activated protein kinase 1 signaling in the treated cells. Our present study suggests that chemical proteomics can be a very useful and powerful technique for identifying the target proteins of small bioactive molecules.
Single-strand DNA Binding of Actinomycin D with a Chromophore 2-Amino to 2-Hydroxyl Substitution
Yoo, Hoon ; Rill, Randolph L. ;
BMB Reports , volume 36, issue 3, 2003, Pages 305~311
DOI : 10.5483/BMBRep.2003.36.3.305
A modified actinomycin D was prepared with a hydroxyl group that replaced the amino group at the chromophore 2-position, a substitution known to strongly reduce affinity for double-stranded DNA. Interactions of the modified drug on single-stranded DNAs of the defined sequence were investigated. Competition assays showed that 2-hydroxyactinomycin D has low affinity for two oligonucleotides that have high affinities (
oligomer) for 7-aminoactinomycin D and actinomycin D. Primer extension inhibition assays performed on several single-stranded DNA templates totaling around 1000 nt in length detected a single high affinity site for 2-hydroxyactinomycin D, while many high affinity binding sites of unmodified actinomycin D were found on the same templates. The sequence selectivity of 2-hydroxyactinomycin D binding is unusually high and approximates the selectivity of restriction endonucleases. Binding appears to require a complex structure, including residues well removed from the polymerase pause site.
The Effects of a High-fat or High-sucrose Diet on Serum Lipid Profiles, Hepatic Acyl-CoA Synthetase, Carnitine Palmitoyltransferase-I, and the Acetyl-CoA Carboxylase mRNA Levels in Rats
Ryu, Mi-Hyun ; Cha, Youn-Soo ;
BMB Reports , volume 36, issue 3, 2003, Pages 312~318
DOI : 10.5483/BMBRep.2003.36.3.312
The purpose of this study was to investigate the effects of altering relative intakes of fat and carbohydrates on serum lipid profiles, hepatic acyl-CoA synthetase (ACS), carnitine palmitoyltransferase-I (CPT-I), and the acetyl-CoA carboxlyase (ACC) mRNA level in Sprague-Dawley rats. For four weeks the rats were fed either an AIN-76 diet or one of its modified diets that were supplemented with 20% beef tallow (high-fat diet, HF) and 66.3% sucrose (highsucrose diet, HS). The HS group had significantly higher serum triglyceride and total cholesterol concentrations when compared with the other groups. Serum LDL-cholesterol concentrations in the HS and HF groups were significantly higher when compared to the normal diet (ND) group. Serum HDL-cholesterol levels of the ND and HS groups were significantly higher than those of the HF group. The hepatic total lipid level of the HF group was significantly higher than those of other groups; triglyceride levels of the HS and HF groups were significantly higher than those of the ND group. Hepatic ACS mRNA levels of the HF group were significantly higher than those of the ND group. Hepatic CPT-I mRNA levels were higher in the HF group than other groups. Also, ACC mRNA levels in the liver increased in the HF group. In conclusion, changes in the composition of dietary fat and carbohydrates could affect the hepatic ACS, CPT-I, and ACC mRNA levels. These results facilitate our understanding of the coordinated regulation of the ACS, CPT-I, and ACC mRNA levels and will serve to enhance our understanding of the molecular mechanisms that underlie the regulation of fatty acid metabolism.
Stimulation of γ-Aminobutyric Acid Synthesis Activity in Brown Rice by a Chitosan/Glutamic Acid Germination Solution and Calcium/Calmodulin
Oh, Suk-Heung ;
BMB Reports , volume 36, issue 3, 2003, Pages 319~325
DOI : 10.5483/BMBRep.2003.36.3.319
Changes in the concentrations of
-aminobutyric acid (GABA), soluble calcium ions, glutamic acid, and the activity of glutamate decarboxylase (GAD) were investigated in non-germinated vs. germinated brown rice. Brown rice was germinated for 72 h by applying each of the following solutions: (1) distilled water, (2) 5 mM lactic acid, (3) 50 ppm chitosan in 5 mM lactic acid, (4) 5 mM glutamic acid, and (5) 50 ppm chitosan in 5 mM glutamic acid. GABA concentrations were enhanced in all of the germinated brown rice when compared to the non-germinated brown rice. The GABA concentration was highest in the chitosan/glutamic acid that germinated brown rice at 2,011 nmol/g fresh weight, which was 13 times higher than the GABA concentration in the non-germinated brown rice at 154 nmol/g fresh weight. The concentrations of glutamic acid were significantly decreased in all of the germinated rice, regardless of the germination solution. Soluble calcium and GAD were higher in the germinated brown rice with the chitosan/glutamic acid solution when compared to the rice that was germinated in the other solutions. GAD that was partially purified from germinated brown rice was stimulated about 3.6-fold by the addition of calmodulin in the presence of calcium. These data show that the germination of brown rice in a chitosan/glutamic acid solution can significantly increase GABA synthesis activity and the concentration of GABA.
Regulation of the Gene Encoding Glutathione Synthetase from the Fission Yeast
Kim, Su-Jung ; Shin, Youn-Hee ; Kim, Kyung-Hoon ; Park, Eun-Hee ; Sa, Jae-Hoon ; Lim, Chang-Jin ;
BMB Reports , volume 36, issue 3, 2003, Pages 326~331
DOI : 10.5483/BMBRep.2003.36.3.326
The fission yeast cells that contained the cloned glutathione synthetase (GS) gene showed 1.4-fold higher glutathione (GSB) content and 1.9-fold higher GS activity than the cells without the cloned GS gene. Interestingly,
-glutamylcysteine synthetase activity increased 2.1-fold in the S. pombe cells that contained the cloned GS gene. The S. pombe cells that harbored the multi copy-number plasmid pRGS49 (containing the cloned GS gene) showed a higher level of survival on solid media with cadmium chloride (1 mM) or mercuric chloride (
) than the cells that harbored the YEp357R vector. The 506 bp upstream sequence from the translational initiation point and N-terminal8 amino acid-coding region were fused into the promoteriess
-galactosidase gene of the shuttle vector YEp367R to generate the fusion plasmid pUGS39. Synthesis of
-galactosidase from the fusion plasmid pUGS39 was significantly enhanced by cadmium chloride and NO-generating S-nitroso-N-acetylpenicillamine (SNAP) and sodium nitroprusside (SN). It was also induced by L-buthionine-(S,R)-sulfoximine, a specific inhibitor of
-glutamylcysteine synthetase (GCS). We also found that the expression of the S. pombe GS gene is regulated by the Atf1-Spc1-Wis1 signal pathway.
Microplate Assay Measurement of Cytochrome P450-Carbon Monoxide Complexes
Choi, Suk-Jung ; Kim, Mi-Ra ; Kim, Sung-Il ; Jeon, Joong-Kyun ;
BMB Reports , volume 36, issue 3, 2003, Pages 332~335
DOI : 10.5483/BMBRep.2003.36.3.332
Cytochrome P450 in microsomes can be quantitated using the characteristic 450 nm absorption peak of the CO adduct of reduced cytochrome P450. We developed a simple microplate assay method that is superior to previous methods. Our method is less laborious, suitable for analyzing many samples, and less sensitive to sample aggregation. Microsome samples in microplate wells were incubated in a CO chamber rather than bubbled with CO gas, and then reduced with sodium hydrosulfite solution. This modification allowed a reliable and reproducible assay by effectively eliminating variations between estimations.