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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Journal DOI :
Korean Society for Biochemistry and Molecular Biology
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Volume & Issues
Volume 30, Issue 6 - Nov 1997
Volume 30, Issue 5 - Sep 1997
Volume 30, Issue 4 - Jul 1997
Volume 30, Issue 3 - May 1997
Volume 30, Issue 2 - Mar 1997
Volume 30, Issue 1 - Jan 1997
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Effect of Trichloroethylene on the Induction of Rat Liver Microsomal Enzymes
Chang, Sung-Keun ; Jeong, Hyo-Seok ; Chai, Se-Ok ; Kim, Ki-Woong ; Park, Sang-Shin ;
BMB Reports , volume 30, issue 4, 1997, Pages 237~239
The effects of trichloroethylene (TRI) on the induction of cytochrome P-450 (CYP) and several other related enzymes in Sprague Dawley rats were investigated Rats were treated with TRI 150. 300. 600 mg/kg body weight in corn oil intra peritoneally once a day for 2 days. The total contents of microsomal CYP and cytochrome
decreased with the increase of TRI concentration. but the activity of p-nitrophenol hydroxylase increased with the increase of TRI dosage (p<0.05). Western blot analysis which utilized monoclonal antibodies against CYP2E1 also showed a significant increase in the CYP2E band density. The increase of the activity of pentoxyresolufin-O-deethylase also was observed with the TRI treatment (p<0.05) although there was no significant increase in the cytochrome CYP2B1/2 in Western blotting The TRI did not affect the induction of aryl hydrocarbon hydroxylase. These findings suggest that the CYP2E1 is the primary enzyme which could be induced by TRI treatment in rats.
Effect of Ganglioside
on the Erythrocyte Glucose Transporter (GLUT1): Conformational Changes Measured by Steady-State and Time-Resolved Fluorescence Spectroscopy
Yoon, Hae-Jung ; Lee, Min-Yung ; Jhon, GiI-Ja ;
BMB Reports , volume 30, issue 4, 1997, Pages 240~245
Interactions between ganglioside
and glucose transporter, GLUT1 were studied by measuring the effect of
on steady-state and time-resolved fluorescence of purified GLUT1 in synthetic lipids and on the 3-O-methylglucose uptake by human erythrocytes. The intrinsic tryptophan fluorescence showed a GLUT 1 emission maximum of 335 nm, and increased in the presence of
by 12% without shifting the emission maximum, The fluorescence lifetimes of intrinsic tryptophan on GLUT1 consisted of a long component of 7.8 ns and a short component of 2,3 ns and
increased both lifetime components. Lifetime components were quenched by acrylamide and KI. Acrylarnide-mduced quenching of long-lifetime components was partly recovered by
However. KI-induccd quenching of short- and long-lifetime components was not rescued by
. The anisotropy of 1.6-diphenyl-1.3.5-hexatriene (DPH)-probed dimyristoylphosphatidylcholine (DMPC) model membrane was also increased with
incorporation, The transport rate of 3-O-methylglucose increased by 20% with
incorporation on the erythrocytes, Therefore,
altered the environment of lipid membrane and induced the conformational change of GLUT1.
CoA Transferase and Malonyl-CoA Decarboxylase Activity of Malonate Decarboxylase from Acinetobacter calcoaceticus
Byun, Hye-Sin ; Kim, Yu-Sam ;
BMB Reports , volume 30, issue 4, 1997, Pages 246~252
Malonate decarboxylase from Acinetobacter calcoaceticus is shown to have malonyl-CoA: acetate CoA transferase. acetyl-CoA: malonate CoA transferase, and malonyl-CoA decarboxylase activity. These enzyme activities were elucidated by isotope exchange reactions. The enzyme modified by N-ethylmaleimide completely lost its malonate decarboxylase activity, whereas it still kept CoA transferases and malonyl-CoA decarboxylase activities. The existence of CoA transferases and malonyl-CoA decarboxylase activity is clear, but their physiological significance is obscure. The catalytic reactions for two eoA transfers and malonyl-CoA decarboxylation proceed via a cyclic mechanism, which is through two covalent intermediates, enzyme-Smalonyl and enzyme-S-acetyL proposed for malonate decarboxylation of the enzyme.
Comparison between Positive and Negative Ion Mode FAB CAD MS/MS Spectra of Linkage-Isomeric Oligosaccharides
Yoo, Eun-Sun ;
BMB Reports , volume 30, issue 4, 1997, Pages 253~257
Negative ion fast atom bombardment (FAB) mass spectra were found to allow the determination of the linkage positions in a series of underivatized linkage-isomeric oligosaccharides. A previous work (Laine et al., 1988) reported that ion patterns of linkage-isomeric trisaccharides could be distinguished by a positive ion. Negative ion FAB collison-activated dissociation (CAD) mass spectrometry (MS) spectra of trisaccharides exhibited better sensitivity than the positive ion mode and provided specific fragmentation patterns according to the linkage positions. Especially, the fragmentations, m/z 205 in F6 and m/z 221 in G6, not occuring in 1-3 or 1-4 linkage. were an indication of 1-6 linkage, by changing collision energies from + 10 eV to +60 eV. The survival ratios of molecular ions in each collision energy set gave support to previous results in which the order of bond stability was 1-6>1-4>1-3 linkage.
Improved Fluorometric Assay Method for Ribonuclease Activity
Lee, Jong-Soo ; Choi, Jong-Soo ;
BMB Reports , volume 30, issue 4, 1997, Pages 258~261
A simple quantitative assay method for ribonuclease activity has been developed. This method is based on the decrease of fluorescence intensity emitted by the ethidium bromide bound to RNA due to the degradation of RNA by ribonuclease. The substrate RNA was reacted with ribonuclease A and the fluorescence intensity was measured after the addition of ethidium bromide. The intensity difference was calculated using a blank reaction mixture containing no RNase. Whole cellular RNA substrate produced a significant error and was not suitable for this assay method possibly because of local microheterogeniety caused by high molecular weight rRNA. but satisfying results were obtained with tRNA substrate. The intensity difference increased linearly by raising enzyme concentration up to
Kunitz Units of ribonuclease A. More refined and reliable results were obtained by use of initial reaction velocities which were calculated from the plots of intensity difference vs time. A linear relationship between initial velocities and enzyme concentrations was observed up to 0.01 Kunitz Units of enzyme.
Purification and Characterization of a Thermostable Alkaline Phosphatase Produced by Thermus caldophilus GK24
Kim, You-Jin ; Park, Tae-Shin ; Kim, Hyun-Kyu ; Kwon, Suk-Tae ;
BMB Reports , volume 30, issue 4, 1997, Pages 262~268
The thermophilic and thermostable alkaline phosphatase was purified to near homogeneity from the osmotic lysis of Thermus caldophilus GK24, The purified enzyme had an apparent molecular mass of 108, 000 Da and consisted of two subunits of 54,000 Da. lsoelectric-focusing analysis of the purified enzyme showed a pi of 7.3. The enzyme contained two Cys residues, and its amino acids composition was quite different from that of Thermus aquaticus YT-1 alkaline phosphatase and Escherichia coli alkaline phosphatase, The optimum pH and temperature of the enzyme were 11.0-11.5 and
respectively. The enzyme was stable in the pH range of 9.0-12.0 at
for 36 h. and the half-life at
(pH 11.0) was 6 h. The enzyme was activated by
and inhibited by EDTA. With
as the substrate, the enzyme had a Michaelis constant
, The enzyme preferentially hydrolyzed the phosphomonoester bond of AMP in ribonucleotides and glycerophosphate.
The Action of Hepatitis B Virus Enhancer 2-Core Gene Promoter in Non-Viral and Retroviral Vectors for Hepatocyte-Specific Expression
Rih, Jeong-Keun ; Oh, Sang-Taek ; Hwang, Deog-Su ; Kim, Sun-Young ; Yim, Jeong-Bin ;
BMB Reports , volume 30, issue 4, 1997, Pages 269~273
Heptocvte-specific expression induced by Hepatitis B virus (HBV) enhancer 2-core gene promoter was examined in various hepatocyte and non-hepatocyte cell lines. using non-viral and retroviral vector systems in which chloramphenicol acetyltransferase (CAT) is used as a reporter. The non-viral plasmid containing the HBV enhancer 2-core promoter exhibited 22 and 66% of CAT activities in hepatoma cell lines. HepG2 and Hep3B, respectively when compared with CAT activity expressed by CMV promoter. The CAT activities, however. were found to be marginal in other tested hepatoma cell lines as well as mouse primary hepatocytes and non-hepatocytes. The HBV enhancer 2 located upstream the CMV promoter did not affect the CMV promoter activity nor provided hepatocyte-specific expression. Transfection of retroviral plasmid DNA containing the HBV enhancer 2-core promoter as an internal promoter exhibited high and specific CAT expression in HepG2 and Hep3B cell lines but the activity value was 5 to 10 fold lower than the non-viral plasmid with identical promoter. These results suggest that the usage of HBV enhancer 2-core promoter for liver specific expression is limited to certain vectors and hepatocyte cell lines.
Purification and Characterization of Acetolactate Synthase from Barley
Chong, Chom-Kyu ; Chang, Soo-Ik ; Choi, Jung-Do ;
BMB Reports , volume 30, issue 4, 1997, Pages 274~279
Acetolactate synthase (ALS) catalyzes the first common step in the biosynthesis of branchedchain amino acids, valine, leucine, and isoleucine. ALS is the target site for several structually diverse classes of herbicides including sulfonylureas, imidazolinones. and triazolopyrimidines. We have purified ALS from etiolated barley shoots to homogeneity. The five major purification steps are ammonium sulfate fractionation, DEAE anion exchange, hydroxylapatite, Bio-Gel A gel filtration, and low pressure Mono-Q chrornatoqraphy. Approximately 170-fold purification was achieved and the yield was 0.45% of initial activity in the crude extract. Both SDS-PAGE and Western blot analysis showed a single polypeptide of ALS with an apparent molecular mass of 64 kDa. The result of nondenaturing gel electrophoresis with activity staining indicated that the molecular mass of its native form is approximately 225 to 250 kDa. The values of
for pyruvate. pl. and optimum pH of ALS were determined to be 2.0 mM, 5.2. and 7.0. respectively Feedback inhibition studies showed that ALS is more susceptible to leucine than valine. And
value of Cadre, a class of irnidazolinones, is about
Chemical Modification Studies of Yeast Farnesyl Protein Transferase
Sohn, Seung-Wan ; Jun, Gyo ; Yang, Chul-Hak ;
BMB Reports , volume 30, issue 4, 1997, Pages 280~284
Phenylglyoxal diethyl pyrocarbonate (DEPC), and 1-cyclohexyl-3-[2-morpholinoethyl]-carbodiimide metho-p-toluenesulfonate (CMC) are modifying reagents specific for arginine, histidine, and aspartate or glutamate, respectively. They were found to inactivate S. cerevisiae farnesyl protein transferase (FPTase). The peptide substrate protected the enzyme against inactivation by CMC and the other substrate farnesyl pyrophosphate showed protection against inactivation by phenylglyoxal. while neither of the two substrates protected the enzyme against DEPC inactivation. These results suggest the presence of aspartate/glutamate, arginine and histidine residues at the active site of this enzyme.
Involvement of Cytochrome c Oxidase Subunit I Gene during Neuronal Differentiation of PC12 Cells
Kang, Hyo-Jung ; Chung, Jun-Mo ; Lee, See-Woo ;
BMB Reports , volume 30, issue 4, 1997, Pages 285~291
It is becoming increasingly evident that significant changes in gene expression occur during the course of neuronal differentiation. Thus, it should be possible to gain information about the biochemical events by identifying differentially expressed genes in neuronal differentiation The PC12 cell line is a useful model system to investigate the molecular mechanism underlying neuronal differentiation and has been used extensively for the study of the molecular events that underlie the biological actions of nerve growth factor (NGF). In this study, we report an application of the recently described mRNA differential display method to analyze differential gene expression during neuronal differentiation. Using this technique, we have identified several cDNA tags expressed differentially during neuronal differentiation. Interestingly, one of these clones was cytochrome c oxidase subunit I (COX I) gene. The differential expression of COX I gene was confirmed by Northern blot analysis as well as RT-PCR. Southern blot analysis of the genomic DNA of PC12 cells revealed that COX I is a single gene. Induction of the oxidative enzyme might reflect the energy requirement in neuronal differentiation.
Identification of Genes Induced by Low Temperature in Rice
Choi, Kyong-Hee ; Choi, Hack-Sun ; Lee, Choon-Hwan ; Kwon, Young-Myung ; Rhew, Tae-Hyong ;
BMB Reports , volume 30, issue 4, 1997, Pages 292~295
Exposure of seedling of rice (Oriza sativa cv.Dongin) to cold stress (
, 7day) induced differential gene expression. Differentially expressed polyadenylated RNA induced by low temperature were isolated and identified from the leaves of rice (Oriza sativa cv.Dongin) seedling by using the technique, differential display of reverse transcription through polymerase chain reaction (DDRT-PCR). Four bands of cDNAs were differentially displayed on the PAGE gel through DDRT-PCR, and among them three bands were those of overexpressed genes while one band was of an underexpressed gene One of the overexpressed cDNA was characterized. The size of the DDRT-PCR product was found to be about 200 bp. The sequence of the cloned DNA was compared with those of GenBank through a BLAST E-Mail server, and it was found to have no homologies in the nucleotide sequence with that of any known DNA: therefore, it was designated as RC101 The expression of the cold-stress induced-gene, RC101, was sustained with Northern Blot analysis by using the cloned DDRT-PCR product as a probe.
Preliminary X-Ray Diffraction Study of Glutathione S-Transferase from Pseudomonas sp. DJ77
Choi, Heung-Soo ; Woo, Ju-Rang ; Lee, Jung-Hee ; Chung, An-Sik ; Ryu, Seong-Eon ; Kim, Young-Chang ; Chung, Yong-Je ;
BMB Reports , volume 30, issue 4, 1997, Pages 296~298
A bacterial glutathione S-transferase from Pseudomonas sp. DJ77 has been crystallized. The crystals diffract to at least
resolution, and belong to the orthorhombic space group
, with cell parameters
. There is one dimer molecule of pGST per crystallographic asymmetric unit. with the crystal volume per protein mass of
and a solvent content of about 47% (v/v).
cDNA Sequences for Asialoglycoprotein Receptor from Human Fetal Liver
Lee, Dong-Gun ; Lee, Sung-Gu ; Kim, Kil-Lyong ; Hahm, Kyung-Soo ;
BMB Reports , volume 30, issue 4, 1997, Pages 299~301
The asialoglycoprotein receptor (ASGPR) was the first described mammalian lectin that mediates the specific binding and internalization of galactose/N-acetylgalactosamine-terminating glycoproteins by hepatic parenchymal cells. H1 and H2 are known as essential subunits of the functional ASGPR. There were close similarities in ASGPR H2 subunits between cultured cell line HepG2 and normal human liver cells including identical sequences at both termini. It was therefore expected that there may be some similarities between the subunits from normal liver cells and fetal liver cells. The two subunits of human fetal liver ASGPR. designated FL-H1 and FL-H2. were cloned from cDNA library by peR and the sequences were compared with the known HI and H2 sequences of HepG2, and the H1 sequence of nornal human liver cells. The results showed that FL-H1 was identical to H1 of HepG2. Whereas FL-H2 contains a 15-bp miniexon, but missing 57-bp at the near upstream from the membrane-spanning domain compared to H2 of HepG2 and normal human liver cells indicating that FL-H2 resulted from a differential splicing compared to HepG2 and normal liver cells.