TPA로 야기된 HL-60 세포의 기질부착 저해작용을 이용한 Protein Kinase C 저해 생약의 탐색

김선희,안종석,김삼용,유관희,안병준 충남대학교 약학대학 의약품개발연구소 1993 藥學論文集 Vol.9 No.-

Thirty-five kinds of herbal drugs, believed to be active for treatment of tumors, were selected as the experimental materials for observing their effects on TPA-induced adherence of HL-60 cell and activity of protein kinase C. They were extracted with ethyl ether(E) and ethyl acetate(EA), methanol(M) in sequence. Among the extracts, Sophorae Flos(EA), Paeoniae Radix(EA), Equisetum hiemale(EA), Phellodendri Cortex(E, EA), Mori Cortex radicis(EA), Ferula assafoetida(E, EA), Sophora subprostrata(EA, M) and Cnidium monnieri(E) inhibited the TPA-in-duced adherence of HL-60 cell more than 50% and Sophorae Flos(E, EA, M), Paeoniae Radix(E, EA, M), Eguisetum hiemale(E), Phellodenri Cortex(E,H), Mori Cortex radicis(M), Ferula assafoetida(M), Sophora subprostrata(EA, M), Cnidium monnieri(M) and Artemisia argyi(M) showed inhibiting effects on the activity of protein kinase C more than 50%. The extracts showing good inhibiting effects on the adherence and enzyme activity were Sophorae Flos(EA), Paeoniae Radix(EA), Phellodendri Cortex(E), Sophora subsprostrate(EA) and Equisetum hiemale(EA).

실험적 뇌지주막하출혈 토끼에서의 Malonate 농도 측정

김재중,하영수,백승렬,장정순,김유삼 대한신경외과학회 1995 Journal of Korean neurosurgical society Vol.24 No.10

Malonate is regarded as a reversible competitive inhibitor of succinate dehydrogenase and malate transport in the Krebs cycle and showed neurotoxicity by persistent NMDA receptor activation due to inhibition of ATP production and glutamate utilization. However, little was known about its biological effects and the range of normal concentration of malonate in central nervous system. In order to understand the relationship between malonate and vasospsasm, malonate concentrations in rabbit model of experimental subarachnoid hemorrhage were measured at 0, 4th, and 7th day following SAH in serum, cerebrospinal fluid, and urine using malonyl-CoA synthetase. The results were as follows : 1) Malonate level is increased significantly in serum at the 4th day after subarachnoid hemorrhage that followed by vasospasm(p<0.01). 2) CSF malonate concentration tends to increase at post-SAH 7th day but statistically not significant. 3) The change of urine malonate concentration is not significant. These results suggest that early increase of serum malonate level is significant because clinically important vasospam begin from the fourth through the seven day after hemorrhage. The increased level of serum malonate at this time is due to impairment of cellular metabolism following delayed cerebral ischemia and may influence to development of vasospasm. In conclusion, the measurement of serum malonate concentration after subarachnoid hemorrhage is one of possible candidates for the early diagnosis of vasospasm.

진균성 부비동염 환자의 상악동내 농성 분비물에 대한 세균 배양 및 항생제 감수성 결과

이윤식,김용재,김재호,정유삼,이봉재 대한이비인후과학회 2000 대한이비인후과학회지 두경부외과학 Vol.43 No.10

The Effect of ZD 1839 (Iressa) in the Treatment of RefractoryNon Small Cell Lung Cancer

Yong Tai Kim,Chul Kim,Joo Hyuk Sohn,So Young Park,Soo Young Park,Nae Choon Yu,Young Sam Kim,Se Kyu Kim,Joon Chang,Kil Dong Kim,Kyung Young Chung,Joo Hang Kim 대한암학회 2003 Cancer Research and Treatment Vol.35 No.6

Purpose: The aim of this study was to evaluate theefficacy and the safety of ZD 1839 (Iressa) as a 3rd or4th line chemotherapy regimen in NSCLC patients whoare refractory to a previous chemotherapy regimen.Materials and Methods: Twenty-five patients who wererefractory to previous chemotherapy were selected forthis study. The eligible patients had an ECOG performancestatus of 0 to 2, and an appropriate end organfunction. ZD 1839 (Iressa) 250 mg/d was orally administereduntil the patients experienced disease progressionor unacceptable toxicity.Results: Twenty-five patients were analyzed. The medianage of the patients was 57 years. The response ratewas 12.0% with partial responses in 3 patients. Fourteenpatients (56%) remained in the stable disease state and8 patients progressed. The median overall survival was9.0 months (95% CI 6.7~11.2). The median progressionfree survival was 3 months (95% CI 2.2~3.8). Hematologicaltoxicities of grade 3 or 4 neutropenia, anemia andthrombocytopenia were absent. Non-hematological toxicitieswere grade 2 or 3 skin rashes in 10 (40.0%) patientsand 1 (4.0%) patient and grade 3 nausea in 3 (12.0%) patients.No patient failed to continue chemotherapy due toany drug-related adverse events.Conclusion: The results suggest that ZD 1839 (Iressa)monotherapy is effective and tolerable as a 3rd or 4th linesalvage treatment for NSCLC patients refractory to previouschemotherapy regimens. (Cancer ResTreat. 2003;35:502-506)

Properties of Malonyl-CoA Decarboxylase from Rhizobium trifolii

Kim, Yu Sam,Lee, Dai Woon,An, Jae Hyung,Song, Jong Hee,Lee, Gha Young 생화학분자생물학회 2000 BMB Reports Vol.32 No.4

A novel gene for malonyl-CoA decarboxylase was discovered in the mat operon, which encodes a set of genes involved in the malonate metabolism of Rhizobium trifolii (An and Kim, 1998). The subunit mass determined by SDS-PAGE was 53 kDa, which correspond to the deduced mass from the sequence data. The molecular mass of the native enzyme determined by field flow fractionation was 208 kDa, indicating that R. trifolii malonyl-CoA decarboxylase is homotetrameric. R. trifolii malonyl-CoA decarboxylase converted malonyl-CoA to acetyl-CoA with a specific activity of 100 unit/㎎ protein. Methylmalonyl-CoA was decarboxylated with a specific activity of 0.1 unit/ ㎎ protein. p-Chloromercuribenzoate inhibited this enzyme activity, suggesting that thiol groups) is(are) essential for this enzyme catalysis. Database analysis showed that malonyl-CoA decarboxylase from R. trifolii shared 32.7 % and 28.1 % identity in amino acid sequence with those from goose and human, respectively, and it would be located in the cytoplasm. However, there is no sequence homology between this enzyme and that from Saccharopolyspora erythreus, suggesting that malonyl-CoA decarboxylases from human, goose, and R. trifolii are in the same class, whereas that from S. erythreus is in a different class or even a different enzyme, methylmalonyl-CoA decarboxylase. According to the homology analysis, Cys-214 among three cysteine residues in the enzyme was found in the homologous region, suggesting that the cysteine was located at or near the active site and plays a critical role in catalysis.

Identification of Amino Acid Residues in the Carboxyl Terminus Required for Malonate - Responsive Transcriptional Regulation of MatR in Rhizobium leguminosarum bv. trifolii

Kim, Yu Sam,Lee, Hwan Young 생화학분자생물학회 1970 BMB Reports Vol.34 No.4

MatR in Rhizobium trifolii is a malonate-responsive transcription factor that regulates the expression of genes, matABC, enabling decarboxylation of malonyl-CoA into acetyl-CoA, synthesis of malonyl-CoA from malonate and CoA, and malonate transport. According to an analysis of the amino acid sequence homology, MatR belongs to the GntR family The proteins of this family have two-domain folds, the N-terminal helix-turn-helix DNA-binding domain and the C-terminal ligand-binding domain. In order to End the malonate binding site and amino acid residues that interact with RNA polymerase, a sitedirected mutagenesis was performed. Analysis of the mutant MatR suggests that Arg-160 might be involved in malonate binding, whereas Arg-102 and Arg-174 are critical for the repression activity by interacting with RNA polymerase.

Properties of Acetyl-CoA Synthetase from Pseudomonas fluorescens

Kim, Yu-Sam,An, Jae-Hyung,Yang, Bu-Hyun,Kim, Kyu-Wan Korean Society for Biochemistry and Molecular Biol 1996 Journal of biochemistry and molecular biology Vol.29 No.4

In Pseudomonas fluorescens grown on malonate as sole carbon source, acetyl-CoA synthetase was induced, suggesting that malonate is metabolized through acetate and then acetyl-CoA. Acetyl-CoA synthetase was purified 18.6-fold in 4 steps to apparent homogeneity. The native molecular mass of the enzyme estimated by a native acrylamide gel electrophoresis was 130 kDa. The enzyme was composed of two identical subunits with a molecular mass of 67 kDa. Optimum pH was 70. The acetyl-CoA synthetase showed typical Michaelis-Menten kinetics for the substrates, acetate, ATP and CoA, whose $K_m$ values were calculated to be 33.4, 74.8, and 40.7 mM respectively. Propionate. butyrate and pentanoate were also used as substrates by the enzyme, but the rate of the formation of the CoA derivatives was decreased in the order of the increase in carbon number. The enzyme was inhibited by the group-specific reagents diethylpyro-carbonate, 2,3-butanedione, pyridoxal-5'-phosphate and N-bromosuccinimide. In the presence of substrates the inactivation rate of the enzyme, by all of the group-specific reagents mentioned above decreased, indicating the presence of catalytically essential histidine, arginine, lysine and tryptophan residues at or near the active site. Preincubation of the enzyme with ATP, $Mg^{2+}$ resulted in the increase of its susceptibility to diethylpyrocarbonate, suggesting that ATP, $Mg^{2+}$ may induce a conformational change in the active site exposing the essential histidine residue to diethylpyrocarbonate. The enzyme was acetylated in the presence of acetyl-CoA, indicating that this is one of acyl-enzyme.

Malonyl - CoA Synthetase from Rhizobium trifolii Purification , Properties , and the Immunological with Those from Bradyrhizobium Japonicum and Pseudomonas fluorescens

Yu Sam Kim,Sun Jong Kwon,Sang Won Kang 생화학분자생물학회 1993 BMB Reports Vol.26 No.2

Isolation of A Novel Malonamidase from Bradyrhizobium Japonicum

Yu Sam Kim,Jung Won Park,Sang Won Kang 생화학분자생물학회 1992 BMB Reports Vol.25 No.8

$quot;Malonamate shuttle$quot; as a model of nitrogen flow by malonamate in Bradyrhizobium japonicum-soybean symbiosis has been proposed. One of the key enzymes of the shuttle system, malonamidase Ela, was purified in an electrophoretically homogeneous form as a constitutive protein from free living Bradyrhizobium japonicum USDA110. The molecular size of the enzyme was 129,000 dalton composed of two identical 59,000 dalton subunits. pI of the enzyme was 5.4. The enzyme was highly specific for malonamate and malonate as substrates and no cofactors or external ions were required for its catalysis. K_m for malonamate and for malonate were 4.2 and 12.5 mM, respectively. Optimal pH for malonohydroxamate formation from malonamate and NH₂OH and for that from malonate and NH₂OH were about 8.0 and 6.5, respectively.

Malonate Metabolism : Biochemistry, Molecular Biology, Physiology, and Industrial Application

Kim, Yu Sam 한국생화학분자생물학회 2002 BMB Reports Vol.35 No.5

Malonate is a three-carbon dicarboxylic acid. It is well known as a competitive inhibitor of succinate dehydrogenase. It occurs naturally in biological systems, such as legumes and developing rat brains, which indicates that it may play an important role in symbiotic nitrogen metabolism and brain development. Recently, enzymes that are related to malonate metabolism were discovered and characterized. The genes that encode the enzymes were isolated, and the regulation of their expression was also studied. The mutant bacteria, in which the malonatemetabolizing gene was deleted, lost its primary function, symbiosis, between Rhizobium leguminosarium by trifolii and clover. This suggests that malonate metabolism is essential in symbiotic nitrogen metabolism, at least in clover nodules. In addition to these, the genes matB and matC have been successfully used for generation of the industrial strain of Streptomyces for the production of antibiotics.