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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
Selecting the target year
Effects of Individual Fatty Acids on Receptor-Mediated Binding, Internalization and Degradation of
Choue, Ryo-Won ; Cho, Byung-Hee Simon ;
BMB Reports , volume 30, issue 1, 1997, Pages 1~6
The ability of Hep-G2 cells to process
under basal conditions was investigated. The receptor-binding and internalization of
increased with the time of incubation in a saturable manner. After 4 h of incubation, 31.4 ng of
was cell bound. The cells rapidly internalized
via specific, receptor-mediated endocytosis. The amount of internalized
reached a maximun of 96.7 ng at 2 h of incubation and remained constant for the next 2 h. The rate of degradation of internalized
proceeded in a linear manner over the entire 4 h of incubation after an initial lag period. The effects of individial fatty acids (C18:0. C18:1, C18:2. and C18:3), differing in their degree of unsaturation. on the receptor-binding, internalization and degradation of
were also investigated. Inclusion of 1.0 mM of each fatty acid into the culture medium significantly increased
metabolism in Hep-G2 cells. Among the fatty acids tested, stearic acid had the least effect on the receptor-binding activity. There were no significant differences among the unsaturated fatty acids in LDL-receptor binding. The effect of individual fatty acids on the
uptake was similar to that of the receptor-binding. showing a significantly lower effect with stearic acid. The amount of degraded material of internalized
was the lowest with stearic acid when it was compared with unsaturated fatty acids.
Kinetic Study of the Lipase-Catalyzed Interesterification of Triolein and Stearic Acid in Nonpolar Media
Chi, Young-Min ;
BMB Reports , volume 30, issue 1, 1997, Pages 7~12
The kinetics of the interesterification of triolein and stearic acid catalyzed by immobilized Rhizopus delemar lipase were studied in a batch operation. In order to clarify the mechanisms of this reaction, three models are discussed under various conditions in terms of the ratio of triolein and stearic acid. The rate constants involved in the proposed model were determined by combining the numerical Gauss-elemination method, and the trial-and-error method so as to fit the calculated results with the experimental data. The accuracy of the obtained rate constants was confirmed after they were substituted for simultaneous differential equations and the equations simulated using an adaptive step-size Runge-Kutta method. Finally, the model which agrees with the calculated results and the experimental data was selected.
Effect of Pyrimidylsalicylate on the Valine Sensitive Acetolactate Synthase Purified from Serroatia marcescens
Yang, Jeong-Hee ; Kim, Soung-Soo ;
BMB Reports , volume 30, issue 1, 1997, Pages 13~17
The inhibitory effect of herbicides such as sulfonylurea derivatives, imidazolinones and pyrimidylsalicylate has been examined on the purified valine sensitive acetolactate synthase (ALS) from Serratia marcescens. The concentration of sulfometuron methyl which inhibits 50% of the ALS activity was 2.5 mM. The required concentrations of triasulfuron, primisulfuron methyl and imazaquin for the 50% inhibition of the ALS activity were 1 mM. The resistance of Serratia ALS to sulfometuron methyl, imazapyr and imazaquin is similar to that of E. coli ALS 1. However, pyrimidylsalicylate showed a potent inhibitory effect on the Serratia ALS almost 13 times more potent than on E. coli ALS II, which is known as herbicide-sensitive isozyme. The inhibitory mode was competitive against pyruvate. 150 value was determined to be
in an assay mixture containing 20 mM pyruvate, and the
, value was calculated to be
from the modified double reciprocal plot of 1/V versus
Repetitive Homologous Sequences in Flanking Region of Gametophytic Self-Incompatibility Allele in Lycopersicon peruvianum
Chung, II-Kyung ;
BMB Reports , volume 30, issue 1, 1997, Pages 18~20
Lycopersicon peruvianum shows a gametophytic self-incompatibility (GSI). GSI is controlled by a single locus (S locus) with multiple alleles. S ribonucleases encoded in S alleles cosegregate with their phenotypes of GSI in genetic cross. To understand the genetic role of S allele in L peruvianum, two large genomic fragments isolated previously were analyzed with total genomic DNAs from several tomato lines generated by cross-pollination. Southern blot analysis with the S allele fragments as probes revealed that the flanking region of S allele contained the highly homologous regions. It is speculated that they may play an important role to prevent genetic cross by self-pollination.
The Effect of NaCI Treatment on the Freezing Tolerance and Protein Patterns of Carrot Callus Suspension Culture
Moon, Soon-Ok ; Park, Sook-Hee ; Cho, Bong-Heuy ;
BMB Reports , volume 30, issue 1, 1997, Pages 21~25
The growth. freezing resistance and electrophoretic protein patterns of carrot callus cultures were investigated following treatment with NaCl for various' intervals at 20
. Following 7 day exposure to 250 mM NaCl. freezing tolerance increased, which was measured by 2.3.5-triphenyl tetrazolium chloride (TTC) assay and fresh weight was reduced compared to control cells. Changes of electrophoretic patterns of total and boiling stable proteins were investigated using one or two dimensional gel system. Several proteins with molecular weight of 43 and 21 kDa increased by NaCl treatment. The most prominent change was detected in 21 kDa protein. The steady state level of this protein increased in NaCl treated cells, but decreased in control cells. Twenty one kDa protein was detected only in the NaCl treated cell when boiling stable protein was analyzed. The isoelectric point of 21 kDa protein was identified as 5.7. The timing of increase of 21 kDa protein was correlated to freezing resistance which implied the role of this protein in the induction of freezing resistance of the cell.
Molecular Analysis of HLA-C Using Polymerase Chain Reaction-Sequence Specific Primers
Lee, Kyung-Ok ; Hong, Sung-Hoi ; Kim, Min-Jung ; Park, Taek-Kyu ; Kim, Yoon-Jung ; Lee, Kyu-Pum ;
BMB Reports , volume 30, issue 1, 1997, Pages 26~32
Of all HLA class I molecules, HLA-C gene products are most poorly understood because they express at a low level on the cell surface compared to HLA-A and -B. In order to identify serologically detectable and undetectable HLA-C antigens, we have established a DNA-based tissue typing method for the HLA-C locus by PCR-SSP (polymerase chain reaction-sequence specific primers). Genomic DNA prepared from Iymphoblastoid 21 B-cell lines and 120 Korean individuals by proteinase K digestion and pheno/chloroform extractions have been typed by PCR-SSP (23 primer mixes were used). The PCR-SSP results of control cell lines were discrepant from serology in 1 case among 21 cases: Cw6 which was negative by serology but positive by PCR-SSP (cell line: MANIKA). Twenty four HLA-Cw "blank" antigens among fifty Korean individuals were completely determined by PCR-SSP DNA typing. HLA-Cw*0101 (15.3%), Cw*1401 (12.3%) and Cw*0701 (11.7%) alleles were frequently found in 120 Korean individual samples. In conclusion. the high level of discrimination for HLA-C alleles may prove useful and informative in the study of transplant survival, and identify the importance of allelic differences, not readily detectable by serology, on host and donor compatibility.
Multiple Age-Associated Mitochondrial DNA Deletions in Mouse Brain
Kim, Jin-Sun ; Kim, Min-Jung ; Kwon, In-Sook ; Song, Eun-Sook ;
BMB Reports , volume 30, issue 1, 1997, Pages 33~36
Age-dependent deletion of mitochondrial DNA (mtDNA) was detected in mouse brain using PCR method. The size of the deleted fragment was 0.5 kb, 0.9 kb. 1.7 kb and 4.3 kb in the region between cytochrome b gene and ATPase 6 gene. The deleted fragment was increased gradually from 3-month to 22month Direct repeat sequence flanking the deletion in 0.5 kb PCR product was TAAT.
Deoxyribonucleic Acid Was Responsible for the Anticoagulatory Effect of an Earthworm, Lumbricus rubellus
Paik, Seung-R. ; Woo, Jeong-Im ; Kim, Gyoung-Mi ; Cho, Jin-Mo ; Yu, Kyoung-Hee ; Chang, Chung-Soon ;
BMB Reports , volume 30, issue 1, 1997, Pages 37~40
Earthworm extracts are known for anti-inflammatory, analgesic. antipyretic, and anticancer effects but can also influence blood circulation. It was previously shown that an earthworm, Lumbricus rubelius. contained a water-extractable anticoagulant which was a heat- and acid-stable molecule with hydrophilic property. In order to uncover the biochemical nature of this molecule, the anticoagulant was processed with various hydrolases such as trypsin, DNase, RNase. and lysozome. When the digested samples were analyzed with an in vitro coagulation test measuring activated partial thromboplastin time (APTT) and agarose gel electrophoresis, the anticoagulant proved to be a relatively homogeneous DNA fragment with relative molecular size around 72 base pairs. Interestingly, the activity was further stimulated with a trypsin digestion. RNA. on the other hand, did not prolong the APTT. It was also demonstrated that the DNA accelerated the antithrombin III (AT-III) inhibition of thrombin from
of 0.34 to 0.16 unit determined with S-2238 as a substrate, whereas heparin, a popular anticoagulant. shifted the value to 0.05. Therefore, it is suggested that the DNA could be considered as an alternative antithrombotic agent to heparin, which would exhibits bleeding side effects.
Promoter Structure Which Affects on the Expression of Yeast MGMT Gene
Choe, Soo-Young ;
BMB Reports , volume 30, issue 1, 1997, Pages 41~45
The present study was performed to analyze the molecular mechanism which dictates the transcription regulation of the
-methylguanine-DNA methyltransferase (MGMT) gene in Saccharomyces cerevisiae. Previously we identified one possible upstream repressing sequence (URS) in MGMT promoter by promoter deletion and competition analysis. In this paper we report another regulatory element (UAS: upstream activating sequence. -213 to -136) which affects the transcription activity of MGMT promoter. Gel mobility shift assay and Southwestern blot analysis using UAS probe showed several specific proteins which were able to bind to this sequence.
Processing of an Intracellular Immature Pullulanase to the Mature Form Involves Enzymatic Activation and Stabilization in Alkaliphilic Bacillus sp. S-1
Lee, Moon-Jo ; Kang, Bong-Seok ; Kim, Dong-Soo ; Kim, Yong-Tae ; Kim, Se-Kwon ; Chung, Kang-Hyun ; Kim, Jume-Ki ; Nam, Kyung-Soo ; Lee, Young-Choon ; Kim, Cheorl-Ho ;
BMB Reports , volume 30, issue 1, 1997, Pages 46~54
Alkaliphilic Bacillus sp. S-1 secretes a large amount (approximately 80% of total pullulanase activity) of an extracellular pullulanase (PUL-E). The pullulanase exists in two forms: a precursor form (PUL-I:
180,000), and a processed form (PUL-E:
140,000). Two forms were purified to homogeneity and their properties were compared. PUL-I was different in molecular weight, isoelectric point,
-terminal amino acid sequence, and stabilities over pH and temperature ranges. The catalytic activities of PUL-I were also distinguishable in the
values for various substrates, and in the specific activity for pullulan hydrolysis. PUL-E showed 10-fold higher specific activities than PUL-I. However. PUL-I is immunologically identical to PUL-E, suggesting that PUL-I is initially synthesized and proteolytically processed to the mature form of PUL-E. Processing was inhibited by PMSF, but not by pepstatin, suggesting that some intracellular serine proteases could be responsible for processing of the PUL-I. PUL-I has a different conformational structure for antibody recognition from that of PUL-E. It is also postulated that the translocation of alkaline pullulanase(AP) in the bacterium possibly requires processing of the
-terminal region of the AP protein. Processing of the precursor involves a conformational shift. resulting in a mature form. Therefore. precursor processing not only cleaves the signal peptide, but also induces conformational shift. allowing development of active form of the enzyme.
Heat-Shocked Drosophila Kc Cells Have Differential Sensitivity to Translation Inhibitors
Han, Ching-Tack ;
BMB Reports , volume 30, issue 1, 1997, Pages 55~59
The heat shock response is a universal stress response observed in all organisms and cultured cells. The response is regulated at both the transcriptional and translational level. Heat shocked Drosophila melanogaster Kc cells are used as the system for the study of translational regulation. In this system non-heat shock messages are associated with polysome but are not translated in a heat shocked condition. To figure out the change in the translation machinery. the effects of translation elongation inhibitors were tested on Kc cells. The result showed that the sensitivity of translation to these drugs changed in heat shocked cells. The significant changes were the decreased inhibition of heat shock protein synthesis by cycloheximide, emetine. and puromycin. and the increased inhibition of heat shock protein synthesis by verrucarin A. implying that the translation elongation mechanism in heat shocked cells changed.
Expression and Characterization of Recombinant Human Cu,Zn-Superoxide Dismutase in Escherichia coli
Kang, Jung-Hoon ; Choi, Bong-Jin ; Kim, Sung-Moon ;
BMB Reports , volume 30, issue 1, 1997, Pages 60~65
Expression of human Cu.Zn-superoxide dismutase (SOD) with activity comparable to human erythrocyte enzyme was achieved in E. coli B21(DE3) by using the pET-17b expression vector containing a T7 promoter. Recombinant human SOD was found in the cytosol of disrupted bacterial cells and represented > 25% of the total bacterial proteins. The protein produced by the E. coli cells was purified using a combination of ammonium sulfate precipitation, Sephacryl S-100 gel filtration and DEAE-Sephacel ion exchange chromatography. The recombinant Cu,Zn-SOD and human erythrocyte enzyme were compared using dismutation activity, SDS-PAGE and immunoblotting analysis. The mass of the subunits was determined to be 15,809 by using a electrospray mass spectrometer. The copper specific chelator. diethyldithiocarbamate (DOC) reacted with the recombinant Cu,Zn-SOD. At
concentrations of DOC, the dismutation activity was not inhibited for one hour but gradually reduced after one hour. This result suggests that the reaction of DOC with the enzyme occurred in two distinct phases (phase I and phase II). During phase I of this reaction, one DOC reacted with the copper center, with retention of the dismutation activity while the second DOC displaced the copper, with a loss of activity in phase II.
Study on the Specificity Alteration of Mammalian UV Endonuclease III
Lee, Jae-Yung ; Kim, Joon ;
BMB Reports , volume 30, issue 1, 1997, Pages 66~72
A mammalian DNA repair enzyme, UV endonuclease III which also functions as a ribosomal protein S3 (rpS3), was purified from mouse cells and characterized. UV endonuclease III was previously cloned and known to yield a peptide of 32 kDa upon expression in E. coli [Kim et al., (1995) J. Bioi. Chem. 270, 13620-13629]. However, biochemically purified UV endonuclease III, which has a sedimentation coefficient of 3.25, appears to have an additional peptide of 28 kDa. It appears that two bands were derived from one complex, judging from the comparison of the nuclease activity on the native and SDS-gel electrophoreses. UV endonuclease III becomes non-specific upon purification and this phenomenon is more significant in the case of pure fractions of the enzyme. Non-specific activity was not influenced by pH or any salt conditions.
Modification of Hepatic Microsomal Cytochrome P450 2E1 Enzyme by Garlic Powder in Rat Hepatocarcinogenesis
Park, Kyung-Ae ; Choi, Hay-Mie ;
BMB Reports , volume 30, issue 1, 1997, Pages 73~79
This study was designed to investigate the effects of dietary garlic powder on cytochrome P450 enzymes and membrane stability in murine hepatocarcinogenesis initiated by diethylnitrosamine (DEN). Male Sprague-Dawley rats received a single intraperitoneal injection of DEN (200 mg/kg body wt) dissolved in saline. After 2 weeks on a basal diet, animals were fed diets containing 0. 0.5. 2.0. or 5.0% garlic powder for 6 weeks, and were subjected to two-thirds partial hepatectomy. The areas of placental glutathione S-transferase (GST-P) positive foci were inhibited in rats fed with garlic diets. GST-P is the most effective marker for DEN-initiated lesions. Hepatic microsomal lipid peroxidation was significantly decreased in rats fed with 2.0 and 5.0% garlic powder diets compared with that observed in the control animals and hepatic microsomal glucose 6-phosphatase (G6Pase) activity was found to increase significantly in rats fed 0.5 and 2.0% garlic powder diets. Thus as little as 0.5% garlic powder has a positive effect on the stability of hepatic microsomal membranes. p-Nitrophenol hydroxylase (PNPH) activity and the level of cytochrome P450 2E1 protein in the hepatic microsomes from rats fed diets containing 2.0 and 5.0% garlic powder were much lower than those of control microsomes. Rats fed 5.0% garlic powder diets exhibited the lowest P450 2E1 activity and protein levels among groups. Pentoxyresorufin O-dealkylase activity and immunoblot (cytochrome P450 2B1) analyses were not different between groups. However, the levels of cytochrome P450 1A1/2 protein in rats fed 0.5 and 2.0% garlic powder were significantly induced compared to controls. These results suggest that 2.0% garlic powder is effective in inhibiting the areas of GST-P positive foci, modulating certain isoforms of cytochrome P450 enzymes and stabilizing the hepatic microsomal membrane. Thus, the selective modification of cytochrome P450 enzymes and membrane stability by dietary garlic powder may influence areas of GST-P positive foci and chemoprevention of post-initiation of rat hepatocarcinogenesis.
Secretion of Human Angiogenin into Periplasm and Culture Medium with Its Eukaryotic Signal Sequence by Escherichia coli
Jung, Woo-Jung ; Choi, Suk-Jung ;
BMB Reports , volume 30, issue 1, 1997, Pages 80~84
The synthesis and secretion of human angiogenin in E. coli by the natural leader sequence has been studied. We constructed a recombinant plasmid containing human angiogenin cDNA which encompassed all the coding region including leader sequence required for secretion. The recombinant plasmid was introduced into a suitable E. coli host. The angiogenin was detected in the culture medium and periplasm upon the induction of gene expression. The molecular weight of the secreted angiogenin was identical to that of authentic angiogenin purfied from human plasma when estimated by SDS-PAGE and immunoblotting. showing that the natural leader sequence was recognized and processed by the secretion machinery of E. coli. The angiogenin concentration in the culture medium reached a maximum within 2 h when expressed at
with 0.02~2 mM IPTG. In contrast, the expression level increased gradually over time up to 11 h at
with 0.002~2 mM IPTG and at
with 0.002 mM IPTG.