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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 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
Involvement of Protein Tyrosine Kinase in Stimulated Neutrophil Responses by Sodium Fluoride
Chung, Ki-Kwang ; Han, Eun-Sook ; Lee, Chung-Soo ;
BMB Reports , volume 30, issue 2, 1997, Pages 89~94
In this study, during the activation of neutrophil responses by sodium fluoride. involvement of protein tyrosine kinase was studied. Respiratory burst lysosomal enzyme release and elevation of
stimulated by sodium fluoride in neutrophils were inhibited by protein kinase inhibitors, genistein and tyrphostin. The inhibitory effect of genistein and tyrphostin on superoxide and
production was less than that of protein kinase C inhibitors, staurosporine and H-7. Staurosporine and H-7 had little or no effect on the release of myeloperoxidase and acid phosphatase stimulated by sodium fluoride. EGTA and verapamil inhibited the elevation of
evoked by sodium fluoride. The inhibitory effect of staurosporine on the elevation of
was less than that of genistein. Phorbol 12-myristate 13-acetate (PMA)-stimulated superoxide production, which is sensitive to staurosporine, was further enhanced by genistein, whereas the stimulatory action of PMA on myeloperoxidase release was inhibited by genistein. A pretreatment of neutrophils with PMA signifcantly attenuated sodium fluoride-evoked elevation of
These results suggest that protein tyrosine kinase may be involved in the activation process of neutrophil responses due to direct stimulation of guanine nucleotide regulatory proteins. In neutrophil responses, PMA-stimulated neutrophils appear to show a different type of inhibition of protein tyrosine kinase.
Molecular Cloning and Expression of cDNAs Encoding Mouse
2,3-Sialyltransferase (mST3Gal III) and
2,3-Sialyltransferase (mST3GaI IV)
Kim, Kyoung-Sook ; Kim, Cheorl-Ho ; Shin, Deug-Yong ; Lee, Young-Choon ;
BMB Reports , volume 30, issue 2, 1997, Pages 95~100
Two kinds of cDNA encoding mouse
2,3-sialyltransferase (mST3Gal III) and
2,3-sialyltransferase (mST3Gal IV) were isolated from mouse brain cDNA library by means of a PCR-based approach. The cDNA sequences included an open reading frame coding for proteins of 374 and 333 amino acids, respectively, and the primary structure of these enzymes suggested a putative domain structure consisting of four regions, like that in other glycosyltransferases. The deduced amino acid sequences of mST3GaI III and IV showed a 98% and 89% identity with rat ST3GaI III and human ST3Gal IV, respectively. Northern analysis indicated that the expression of mST3Gal III mRNA was abundant in heart, liver and adult brain, while that of mST3GaI IV mRNA was detected in all tissues tested except for testis, but the level was the highest in liver. Soluble forms of mST3GaI III and IV transiently expressed in COS cells exhibited enzyme activity toward acceptor substrates containing the terminal either
1,4GlcNAc sequences. The substrate preferences of both enzymes were stronger for tetrasaccharides than for disaccharides.
Detection and Characterization of Novel Extracellular Phospholipase
in Urine of Patients with Acute Pyelonephritis
Park, Jae-Hyeun ; Lee, Jee-Hye ; Baek, Suk-Hwan ; Moon, Tae-Chul ; Lee, Jong-Myung ; Kim, Nung-Soo ; Nam, Kyung-Soo ; Chang, Hyeun-Wook ;
BMB Reports , volume 30, issue 2, 1997, Pages 101~105
activity has been detected in urine of patients with acute pyelonephritis (APN). This enzyme required micromolar
ion for its maximum activity and showed a broad range of pH (4.5~10) optimum. Urine enzyme hydrolyzed phosphatidylethanolamine (PE) and phosphatidylserine (PS) more effectively than phosphatidylcholine (PC).
activity in the urine of patients with APN was about 5-fold higher than that of healthy individuals. When urine was subjected to heparinSepharose column chromatography, phospholipase
activity was detected in both heparin-non-binding and binding fractions. Both phospholipase
activities were sensitive less than a micromolar calcium concentration and did not react with anti-human 14-kDa group II phospholipase
monoclonal antibody, HP-l. These findings suggest that two kinds of novel extracellular phospholipase
. which may not belong to the 14-kDa group II phospholipase
family, exist in the urine of patients with APN.
Extracellular Matrix of Fresh and Cryopreserved Porcine Aortic Tissues
Shon, Yun-Hee ;
BMB Reports , volume 30, issue 2, 1997, Pages 106~112
The effect of cryopreservation on extracellular matrix was studied with the ultimate objective of permiting a prediction of the tendency of aorta conduit tissue to calcify following transplantation. Cryopreserved and fresh porcine aorta conduit tissues were extracted using guanidine-hydrochloride (Gdn-HCl) followed by sequential digestion of the tissues with collagenase, elastase, and papain. Glycosaminoglycans (GAGs) of the proteoglycans (PGs) were isolated and quantitated. Gdn-HCl extracted about 61% and 62% of the total GAG (proteoqlycan) material from cryopreserved and fresh tissues, respectively. Collagenasesolubilized proteoglycans from Gdn-HCl extracted tissue represented 20% and 13%, respectively, of the total GAGs present in cryopreserved and fresh tissues. Subsequent elastase hydrolysis of collagenase-digested tissue released about 11% of total GAGs from cryopreserved tissue and 16% from fresh tissue. The remaining 8%, from cryopreserved tissue, and 9%, from fresh tissue, of the total GAGs were obtained after using a papain hydrolysis. There was essentially no difference between fresh and cryopreserved tissues in the relative distribution of proteoglycans in the extracts and digestions except in the initial digestion step where more proteoglycans were obtained from collagenase solubilization of cryopreserved tissue than fresh tissue (p<0.05). The histologic status of the fresh and cryopreserved porcine aortic conduit did not differ markedly. The normal tissue architecture was not affected markedly by the cryopreservation procedure as neither alteration of elastic structure, fibrous proteins nor alteration of nuclear distribution or smooth muscle cell morphology was detected. Quantitative tissue mineral studies revealed that the mean calcium content of the cryopreserved aorta conduit tissue
was higher than that of the fresh tissue
. The mean phosphorus content was
wet tissue from cryopreserved tissue and
wet tissue from fresh tissue. The study indicates that there is no significant alteration in the distribution of PGs in properly cryopreserved tissue, but the total calcium level appears to be increased in tissue cryopreserved by the cryopreservation process used in this study.
Chemical Modification of Cysteine Residues in Hafnia alvei Aspartase by NEM and DTNB
Shim, Joon-Bum ; Kim, Jung-Sung ; Yoon, Moon-Young ;
BMB Reports , volume 30, issue 2, 1997, Pages 113~118
Aspartase from Hafnia alvei was inactivated by N-ethylmaleimide (NEM) and 5,5' -Dithiobis-(2-znitrobenzoic acid) (DTNB) following pseudo-first order kinetics. Their apparent reaction orders were 0.83 and 0.50 for NEM and DTNB modifications, respectively, indicating that inactivation was due to a sulfhydryl group in the active site of aspartase and participation of the sulfhydryl group in an essential step in the catalytic reaction. When aspartase was modified by DTNB, the enzyme activity was restored by dithiothreitol treatment, indicating that cysteine residuetsl islarel possibly at or near the active site. The pH-dependence of the inactivation rate by NEM suggested that an amino acid residue having pK value of 8.3 was involved in the inactivation. When aspartase was incubated with NEM and L-aspartate together, L-aspartate markedly protected the enzyme from inactivation by NEM, but the other reagents used did not.
Enhances Tyrosine Phosphorylation of Two Cellular Proteins in HEL Cells
Lim, Chang-Su ; Chun, Jeong-Seon ; Sung, Soo-Kyung ; Lee, Kyu-Cheol ; Lee, Chan-Hee ;
BMB Reports , volume 30, issue 2, 1997, Pages 119~124
is a multifunctional polypeptide that exerts biological roles including cell proliferation, differentiation, extracellular matrix deposition and apoptosis in many different cell types.
, although known as a negative growth regulator, has not been tested in human embryo lung (HEll cells. This study attempts to understand the role of
on growth control of HEL cells in relationship to tyrosine phosphorylation pattern of cellular proteins. In density-arrested HEL cells treated with
, analysis of Western immunoblot showed induction of tyrosine phosphorylation of two major cellular proteins (15 kDa and 45 kDa). In normal proliferating HEL cells with different concentrations of serum, further analysis indicated that the increase in tyrosine phosphorylation of a 45 kDa protein was regulated in serum concentration-dependent manner. However, in proliferating HEL cells treated with
, tyrosine phosphorylation of 45 kDa was down-regulated. Calcium involvement in the regulation of tyrosine phosphorylation of 45 kDa and 15 kDa proteins was also examined. Tyrosine phosphorylation of 15 kDa protein but not of 45 kDa protein was regulated by exogenous calcium. The level of tyrosine phosphorylation of 15 kDa protein was low at reduced caclium concentration and high at elevated caclium concentration.
reversed the pattern of tyrosine phosphorylation of 15 kDa protein. These results suggest that tyrosine phosphorylation of 45 and 15 kDa proteins in HEL cells may be controlled depending on the physiological status of the cells, i.e., low in arrested cells and high in proliferating cells. And the tyrosine phosphorylation of the two proteins appears to be down- or up-regulated by
Catalytic and Structural Properties of Pyridoxal Kinase
Cho, Jung-Jong ; Kim, Se-Kwon ; Kim, Young-Tae ;
BMB Reports , volume 30, issue 2, 1997, Pages 125~131
This work reports studies of the catalytic and structural properties of pyridoxal kinase (ATP: pyridoxal 5' -phosphotransferase, EC. 18.104.22.168), Pyridoxal kinase catalyzes the phosphorylation of vitamin
(pyridoxal, pyridoxamine, pyridoxine) using ATP-Zn as a phosphoryl donor. The enzyme purified from brain tissues is made up of two identical subunits of 40 kDa each. Native enzyme was inhibited by a substrate analogue, pyridoxal-oxime. Limited chymotrypsin digestion of pyridoxal kinase yields two fragments of 24 and 16 kDa with concomitant loss of catalytic activity. These fragments were isolated by DEAE ion exchange chromatography and used for binding studies with fluorescent ATP and pyridoxal analogues. The spectroscopic properties of both fluorescent pyridoxal analogue and Anthraniloyl ATP (Ant-ATP) bound to the 24 kDa fragment are indistinguishable from those of both pyridoxal analogue and Ant-ATP bound to the native pyridoxal kinase, respectively. The small 16 kDa fragment, generated by proteolytic cleavage of the kinase, does not bind any of the substrate analogues. Binding characteristics of Ant-ATP were extensively studied by measuring the changes in fluorescence spectra at various conditions. From the results presented herein, it is postulated that the structural domain associated with catalytic activity comprises approximately one-half of the molecular mass of pyridoxal kinase (24 kDa). whereas the remaining portion (16 kDa) of the enzyme contains a regulatory binding domain.
Subunit Organization of Bacterial Malonate Decarboxylases: The Smallest
Subunit as an Acyl-Carrier Protein
Byun, Hye-Sin ; Kim, Yu-Sam ;
BMB Reports , volume 30, issue 2, 1997, Pages 132~137
In order to compare molecular structure, malonate decarboxylases from Acinetobacter calcoaceticus, Pseudomonas fluorescens, and Pseudomonas putida aerobically grown on malonate, were purified by the method employing streptomycin sulfate treatment, chromatography with PBE 94 and
agarose. Molecular masses were estimated to be 185, 200, and 200 kDa, respectively. All malonate decarboxylases were multimeric enzymes consisting of four different subunits,
. The molecular masses of the Pseudomonas enzyme subunits were
; which are very similar to those,
of Acinetobacter enzyme. The
of the active form of the enzymes was acetylated. The acetyl group may form a thioester bond with the thiol group of the prosthetic group covalently linked to the enzyme. It suggests that such molecular organization is common in all malonate decarboxylases.
Characterization of the Stearic Acid-Induced Uncoupling of Mitochondrial Respiration
Chun, Sun-Bum ; Ho, Sung-Sook ; Han, Sang-Hwa ;
BMB Reports , volume 30, issue 2, 1997, Pages 138~143
In order to assess controversial' proposals concerning the fatty acid-induced uncoupling of mitochondrial oxidative phosphorylation, we investigated the interaction of stearic acid with key mitochondrial proteins and measured the effect of stearic acid on the respiration of cytochrome c oxidase vesicles. Electron paramagnetic resonance spectra of spin-labeled stearic acid clearly demonstrated that cytochrome c oxidase interacts strongly with stearic acid. However, the respiration of detergent-solubilized cytochrome c oxidase was not altered significantly by stearic acid. Surprisingly, adenine nucleotide carrier, which was assumed to bind and translocate fatty acid anions in the Skulachev model of uncoupling, did not bind stearic acid at all. The respiration rate of cytochrome c oxidase vesicles was increased by ~70% in the presence of
stearic acid and this uncoupling was attributed to a simple protonophoric effect of stearic acid.
Preparation of Diphtheria Toxin A Chain from Escherichia coli
Lee, Jong-Soo ; Yoon, Kyoung-Bum ; Park, Jong-Won ; Choi, Suk-Jung ;
BMB Reports , volume 30, issue 2, 1997, Pages 144~149
An expression vector was constructed containing the gene encoding diphtheria toxin A (DTA) which was placed after a T7 promoter. Cytoplasmic expression of the DTA gene resulted in the formation of an insoluble inclusion body. The inclusion body was collected after the complete lysis of the cell, and subsequent washing with 0.1% Triton X-100 released 16~30% of DTA protein from the inclusion body along with other contaminating proteins. The released DTA protein was purified by dialysis. The remaining pellet was dissolved in 8 M urea containing 5%
, and the denatured DTA was renatured by the dilution-dialysis method. The total yield was 35%, and about 5 mg DTA was obtained from 1 L culture. The DTA protein has a free sulfhydryl group exposed to the protein surface, and was shown to have a tendency to dimerize through disulfide formation in the absence of
. The utility of the sulfhydryl group was tested for the construction of recombinant toxins.
Purification and Characterization of Tyrosinase from Solanum melongena
Lee, Jong-Liong ; Kong, Kwang-Hoon ; Cho, Sung-Hye ;
BMB Reports , volume 30, issue 2, 1997, Pages 150~156
Tyrosinase was purified from Solanum melongena by ammonium sulfate precipitation, Sephadex G-150 and DEAE-Sephacel column chromatography. The molecular weight of the purified tyrosinase was approximately 88,600 daltons with 805 amino acid residues. The amino acid composition showed the characteristic high contents of glycine, glutamic acid and serine residues. The enzyme had high substrate specificity towards (+)-catechin. The
, value for L-DOPA was 20.8 mM. L-ascorbic acid,
, sodium diethyldithiocabamate, KCN and
had strong inhibitory effects on enzyme activity. Sodium diethyldithiocabamate was a competitive inhibitor of the enzyme with a
. The optimum pH of the enzyme was 9.0 and the optimum temperature was
with L-DOPA as a substrate. In addition, the activity was enhanced by addition of
, but decreased in the presence of
Characteristics of the Inhibitory Action of Protease Inhibitors on the Glucose-6-phosphate Transporter
Choi, Joon-Sig ; Shin, Jeong-Sook ; Choi, Hong-Sug ; Park, Jong-Sang ;
BMB Reports , volume 30, issue 2, 1997, Pages 157~161
The present paper reports characteristics and specificity of the inhibitory action of
(TPCK) on the glucose6-phosphate transporter of rat liver microsomes. The TLCK-induced inhibition was pH dependent. The inhibition constants for TPCK were determined by following pseudo-Lst order reaction mechanism. The inhibition was protected by preincubation with excess amount of glucose-6-phosphate. The results proved that (a) TLCK inactivates the microsomal glucose-6-phosphate transporter, (b) the inhibition results from the modification of sulfhydryl groups of the transporter.
5' Processing of RNA I in an Escherichia coli Strain Carrying the rnpA49 Mutation
Jung, Young-Hwan ; Park, Jung-Won ; Kim, Se-Mi ; Cho, Bong-Rae ; Lee, Young-Hoon ;
BMB Reports , volume 30, issue 2, 1997, Pages 162~165
RNA I. a negative controller of ColE1-type plasmid replication, is metabolized by several RNases in Escherichia coli. Two small derivatives of RNA I are accumulated at nonpermissive temperatures in an E. coli strain carrying the rnpA49 mutation, a thermosensitive mutation in the rnpA gene encoding the protein component of RNase P. A primer extension analysis was carried out to compare 5' processing of RNA I in the E. coli rnpA49 cells at both permissive and nonpermissive temperatures. Derivatives of RNA I having different 5' ends were observed in the cells grown at permissive and nonpermissive temperatures. Some of the derivatives may be generated by the cleavage of RNase P.