정상 및 Phenylhydrazine으로 처치한 토끼 적혈구의 Adenosine Deaminase Isozymes의 활성도에 관하여

李鉉國,車鍾希,李奉宰,金良均,李根培 조선의대 1984 The Medical Journal of Chosun University Vol.9 No.1

Stuides on Histone ( Ⅵ ) : Histones from Normal and Myelocytic Leukemia Leucocytes

이근배,이희성,전재수,최재득 생화학분자생물학회 1980 BMB Reports Vol.8 No.1

The five major fractions of histone from normal and myelocytic leukemia leucocytes were prepared by the procedure of Johns. Electrophoretic behaviors on acrylamide gels and amino acid composition of various histone fractions from normal and leukema leucocytes: were compared:: The results obtained were as follows 1. The yields of whole hisfome isolated from normal and : inyelocytic leukemia leucocytes recovered were 37.08㎎ and 58.64㎎ per lg packed cells, respectively. 2. A marked increase in the amount of fraction KAS (f2b) from leukemia leucocytes was observed. These events were apparently correlated with a parallel decrease in the fractions KAP (f1) and ARE (f3). The .amount of fraction fZa (GRK+LAK) was almost to those of corresponding fraction of normal leucocytes. 3. Amino acid analysis showed the fraction ARE (f3) from leukemia cells contained markedly greater amounts of e-N-monomethyllysine and e-N-dimethyllysine than that from the normal cells.

肝組織에서의 Glucagon의 Cholesterol合成 抑制機轉에 對한 硏究

李根培,車鐘希,朴悅,高光三 朝鮮大學校 附設 醫學硏究所 1983 The Medical Journal of Chosun University Vol.8 No.1

The inhibitory mechanism of glucagon on hepatic cholesterol biosynthesis in rat was studied by the measurement of incorporation of tritium from ^(3)H₂O into cholesterol, and intermediary metabolite of lipids. But the rate of cholesterol synthesis was not affected. The hepatic acetyl-CoA carboxylase activity and malonyl-CoA concentration were decreased to 25~35% of control value during the same period after glucagon injection. These data indicates that one site of action of glucagon in regulating fat biosynthesis is at the acetyl-CoA carboxylase reaction.

Histone 분획 H1의 Subfraction 에 관한 연구

이경효,이희성,이근배 중앙대학교 의과대학 의과학연구소 1975 中央醫大誌 Vol.1 No.1

The very lysine-rich histones(H1) from normal leucoctyes, myelocytic leukemia leucocytes, human and bovine colostrum, and bakers' yeast were prepared according to the method of Johns, and Phillips and Johns. The purity of H1 fractions from various sources was checked by 15% polyacrylamide disc gel electrophoresis. The amino acid composition of each H1 fraction were determined by ion exchange chromatography procedure described by Spackman et al. These very lysine-rich histones were, further, subfractionated by chromatography on Amberlite IRC-50 with a shallow guanidinium chloride gradient. The results obtained were summarized as follows: 1) The yield of H1 fractions from various sources greatly differed each other. For example, in myelocytic leukemia leucocytes the levels of H1 was many times that of H1 in normal colostrum, suggesting that the levels of H1 is dependent on the function of tissue proper. 2) The electrophoretic mobility of individual very lysine-rich histone fractions gave similar patterns to those of corresponding fraction from various sources. 3) The chromatographic profiles of very lysine-rich histones from leucocytes, colostrum and bakers' yeast revealed that there were significant differences in molecular species and relative amounts between various biological species. 4) Amino acid composition of histone H1 examined in the present studies, could be classified into two categories. One of these groups resemble the very lysine-rich histone found in eukaryotic cells, i.e., normal leucocytes, human and bovine colostrum. The other group, which may be confined to the myelocytic leukemia leucocytes and bakers' yeast, exhibited quite lower Lys/Arg ratio which due to lesser contents of arginine residues in the subfraction molecule.

Ehrlich 복수암 세포의 지질대사에 관한 연구

이희성,하창우,이근배 중앙대학교 의과대학 의과학연구소 1977 中央醫大誌 Vol.2 No.1

We have studied on lipids of Ehrlich ascites tumor cells (EATC). Lipids were extracted by Kennedy and Kanfer(1964) method and lipoproteins were separated by paper electrophoresis. For the identification and assay of lipid extracts, thin-layer chromatography was employed. The incorporation of ^32P-orthophosphate into phospholipids of EATC was also observed. The results obtained were summarized as follows; 1. The total lipid content of EATC was 47.15 ㎎ per g wet weight; of this 32.2㎎ per g was neutral lipid and 14.95 ㎎ per g was phospholipid. 2. The amounts of neutral lipid fractions of EATC, i.e., free cholesterol, esterified cholesterol, mono-, di-, triglyceride and free fatty acid were 2.06㎎, 1.75㎎, 4.71㎎, 7.15㎎, 10.74㎎ and 2.21㎎, respectively. The amount of triglyceride of EATC was similar to those of normal mouse liver. 3. Phospholipid composition of EATC was phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine and unidentified phospholipid, and contents of the individual phospholipids showed that the over-all composition were very similar to normal mouse liver. 4. Lipoprotein fractions of EATC were chylomicron, β-lipoprotein, α-lipoprotein, and prealbumin. Chylomicron fraction contained a larger amount of lipid and α-lipoprotein fraction contained a smaller amount of lipid. 5. The incorporation of ^32P into phospholipids reached a maximum 5 minutes after incubation.

티탄鐵鐄의 鹽素化反應에 關한 硏究

李根培,林炳五 弘益大學校 1980 弘大論叢 Vol.12 No.-

The chlorination of ilmenite in the presence of activated carbon has been investigated in a horizontal silica glass tube with catalyst and non-catalyst. Results are as follows; 1. In both cases the pseudo first order kinetics on ilmenite has been observed with excess activated carbon and chlorine gas at 320∼900℃. 2. The mixture of CaCl₂-NaCl as a catalyst shows a significant effect on increase of the reaction rate. The rate constant for non-catalytic reaction is 7.4×10??(min.??) at 800℃ and that for catalytic reaction is 5.8(min.??) at 500℃. 3. The activation energies for non-catalytic and catalytic reaction are 15.6㎉/mole and 7.1㎉/mole, respectively. 4. A 99.902% TiCl₄ has been separated from the produced mixture of TiCl₄-FeCl₃ by the successive methods of leaching with n-heptane and distillation. But the product from the simple distillation has been found only 99.848% TiCl₄. 5. A further purification of TiCl₄ to 99.998% is carried out by distillation of 99.902% TiCl₄ that was mixed with antimony powder and sodium hydroxide.

실험적 망상적혈구 증다증에 있어서 산화환원 효소활성의 변동 I : Ⅱ. Glucose 6-phosphate dehydrogenase

박주홍,이근배 朝鮮大學校 附設 醫學硏究所 1991 The Medical Journal of Chosun University Vol.15 No.1

토끼에 Phenylhydrazine을 주사한 후 시일이 경과하는 동안에 채혈한 혈액의 적혈구에서 glucose-6-phosphate dehydrogenase(D-glucose-6-phosphate : oxidoreductase, EC 1. 1. 1. 49)의 총 활성도의 변동과 세포질 및 mitochondria에 함유된 glucose-6-phosphate(G-6-P) dehydrogenase의 활성도의 변화를 관찰하여 다음과 같은 결과를 얻었다. G-6-P dehydrogenase의 활성도의 측정은 Komberg 및 Horecker의 방법에 따랐다. 1. 정상흰쥐 적혈구중의 망상적혈구의 수는 약 2.9%였다. 2. 정상 적혈구의 G-6_P dehydrogenase의 활성도는 약 3.8unit/㎖ RBC였으며 세포질 분획에는 약 99.0%가 mitochondria 분획에 약 1.0%가 분포되어 있었다. 3. Phenylhydrazine 주사후 5~6일이 경과하면 망상적혈구의 수는 최고치에 달하는 바 약 35배의 증가를 보였다. 이때 적혈구내의 G-6-P dehydrogenase의 총 활성도도 최고치에 달하는바, 정상치의 약 1.4배의 증가를 보였다. 총 활성도의 약 97.5%가 세포질 분획에, 약 2.5%가 mitochondria 분획에 증명되었다. 4. 망상적혈구증다증의 유발기에 있어서 G-6-P dehydrogenase의 총 활성도의 증가는 주로 세포질 분획내의 이 효소의 증가에 기인하였다.(Ks=0.0208, Kd=0.0042, t1/2=165.0 hours). 5. Mitochondria 분획내의 isozyme도 정상치의 최고 약 22.1배 증가하였다.(Ks=0.0013, Kd=0.0010, t1/2=69.3 hours). 6. 이 효소의 합성 및 분해속도 상수는 세포질 분획에 있어서는, Ks=0.0123, Kd=0.0464, t1/2=56.3 hours이며 mitochondria 분획에 있어서는, Ks=0.0003, Kd=0.0146, t1/2=47.5 hours 이었다. The activity of glucose 6-phosphate dehydrog-enase(D-glucose 6-phosphate : NADP 1-oxidoreductase, EC 1, 1, 1, 49) isozymes in rat red blood cells were studied during phenylhydrazine-induced reticulocytosis and recovery period from it. Glucose 6-phosphate dehydrogenase activity was assayed by the method of Kornberg and Horecker. The results obtained were as follows: 1. Red blood cells of normal rabbit contained about 2.9% of reticulocytes. 2. Total activity of G-6-P dehydrogenase in normal rat erythrocyte was 3.8 units and the relative activity of cytosolic and mitochondrial isozymes were 99.3% and 0.07% respectively. 3. The reticulocyte count in phenylhydrazine-treated rabbit increased to about 38 times on Day 4, Total G-6-P dehydrogenase activity reached maximal on Day 4, showing about 5.1 units which is about 1.3 fold higher than normal value. The increase in the total G-6-P dehydrogenase activity during induction of reticulocytosis was mostly due to the increase in the activity of cytosolic isozyme (Ks= 0.0208 : Kd=0.0042 ; t1/2=165.0 hours), and partly due to the increse in the activity of the mitochondrial isozyme (Ks=0.0013 ; Kd= 0.0100: t1/2=69.3 hours). 4. The rate constants following recovery from phenylhydrazine treatment were: Ks=0.04 64 : Kd=0.0123 ; t1/2=56.3 hours for cytosolic isozyme and Ks=0.0003 : Kd=0.0146 : t1/2=47.5 hours for mitochondrial isozyme.

실험적 망상적혈구 증다증에 있어서 산화환원 효소활성의 변동 : Ⅰ. Sorbitol dehydrogenas

허광덕,이근배 朝鮮大學校 附設 醫學硏究所 1991 The Medical Journal of Chosun University Vol.15 No.1

페닐히드라진을 정용한 토끼에 있어서 망상적혈구가 생성하였다가 소실되는 동안의 솔비톨 데히드로게나의 활성도 변화, 즉 이 효소의 세포질 및 미토콘드리아 분획내의 isozyme의 합성 및 분해속도를 측정하여 음과 같이 결론을 얻었다. 토끼의 정상 적혈구속에 솔비톨 데드로게나제의 활성을 확인하였다. 세포질내에는 적혈구 ㎖당 20.2 단위, 미토콘드라아 분획에는 적혈구 ㎖당 14.3단위의 활성도를 증명하였다. 세포질 분획속의 효성활성도는 페닐히드라진을 적용한지 5일후에 최고에 달하였으며 정상 토끼에 비하여 7.5배의 증가를 보여주었다.(K_(s)=2.7267;K_(d)=0.0184; t 1/2=37.9 hours). 미토콘프라아 분획속의 이 효소의 활성도는 페닐히드라진을 적용한지 역시 5일후에 최고에 달하였으며 상 토끼에 약 5배의 증가를 보여준다. 이 때의 활성도는 세포질 분획속의 활성도의 약 절반에 해당된다.(K_(s)=2.0520;K_(d)=0.0270; t 1/2=25.7hours.) 각 분획속의 이 효소 활성도의 증가 및 감소는 페닐히드라진으로 유발된 reticulocyte의 증가와 소멸해는 경과와 거의 일치한다. 이 현상을 세포분화에 따르는 에너지 요구 및 세포구성 성분의 형성에 관련시켜 비톨의 역할에 대하여 논의하였다. The rate of biosynthesis and degradation of sorbitol dehydrogenase isozymes of red blood cells from phenylhydrazine-treaed rabbits have been studied. The results obtaind were as follows: In normal red blood cells of rabbit, appreciable levels of sorbitol dehydrogenase activity were detected. The activity of sorbitol dehydrogenase in the cytosolic and mitochondrial fractions were increased significantly after phenylhydrazine treatment, and the maximal values were attained at the 5th day, which were about 7.5 times the initial value fot cytosotic isozyme and 5 time for mitochondrial isozyme, respectively. The pattern of time course changes in these two isozymes were quite similar each other. However, the levels of the mitochondrial isozyme was about one-half of that of the cytosolic isozyme. Our experimental data show that Ks value in the mitochondria of reticulocyte during the induction period of reticulocytosis was about 4.6-fold higher than that in the recovery period, while Kd in the induction period was almost equal to that in the recovery period. The increase in total sorbitol dehydrogenase activity during the induction of reticulocytosis appeared to be primarily due to the increase in the cytoslic isozyme (Ks=2.7267: Kd=0.0l84: t 1/2=37.9 hours) and partly due to the increase in the mitochondrial isozyme during reticulocytosis (Ks=2.0520: Kd=0.0270: t 1/2=25.7 hours). The rate constant dyring recovery period from phenylhydrazine treatment were : Ks=0.4311: Kd=0.03808: t 1 /2=22.5 hours for the mitochondrial isozyme and Ks=0.5820: Kd=0.0291: t 1 / 2=23.8 hours for the cytosolic isozyme. Possible role of sorbitol on the energy yielding and formation of cell components during red blood cell maturation was discussed.

실험적 적혈구 증다증에 있어서 전위효소활성의 변동 : Ⅲ. L-Ornithine Aminotransferase

홍영수,이근배 朝鮮大學校 附設 醫學硏究所 1991 The Medical Journal of Chosun University Vol.15 No.1

토끼에 Phenylhydrazine을 주사한 후 시일이 경과하는 동안에 채혈한 혈액의 적혈구에서 ornithine aminotransferase(L-oi-nithine : 2 oxoacid aminotransferase, EC(2.6.1.13)의 총 활성도의 변동과 세포질 및 mitochondria에 함유된 ornithine aminotransferase의 활성도의 변화를 관찰하여 다음과 같은 결과를 얻었다. ornithine aminotransferase의 활성도의 측정은 김형노 및 공동연구자의 방법에 따랐다. 정상토끼 적혈구증의 망상적혈구의 방법에 따랐다. 정상 적혈구의 ornithine aminotransferase의 활성도는 약 14.1unit/㎖ RBC였으며 세포질 분획에는 약 93.6%가 mitochondria분획에 약 2.4%가 분포되어 있었다. Phenylhydrazine후사후 5~6일이 경과하면 망상적혈구의 수는 최고치에 달하는바 약35배의 증가를 보여 준다. 적혈구내의 ornithine aminotransferase의 총 활성도도 최고치에 달하는바, 정상치의 약9.4배의 증가를 보여준다. 총 활성도의 약 97.3%가 세포질 분획에, 약2.7%가 mitochondria 분획에 증명된다. 망상적혈구증다증의 유발기에 있어서 ornithine aminotransferase의 총 활성도의 증가는 주로 세포질 분획내의 이 효소의 증가에 기인하는 것이다. (Ks=0.0339, Kd=0.0102, t1/2=72.2 hours). Mitochondria 분획내의 isozyme도 정상치의 최고 약22.1배 증가한다. (Ks=0.0339, Kd=0.0102, t1/2=67.9 hours) Phenylhydrazine 주사하였을 때 이 효소의 합성 및 분해속도 상수는 세포질 분획에 있어서는, Ks=0.4382, Kd=0.0332 t1/2=20.9 hours이며 mitochondria 분획에 있어서는, Ks=0.0060, Kd=0.0401, t1/2=17.3 hours이다. The activity of ornithine aminotransferase(L-ornithine : 2-oxoacid aminotrasnferase, EC 2. 6. 1. 13) isozymes in red blood cells were studied during induction of and recovery from phenylhy-drazine-produced reticulocytosis is from rabbit. Ornithine aminotransferase activity was assayed by the method of Kim et al. The results obtained were as follow : 1. Red blood of normal rabbit contained about 2.9% of reticulocytes. 2. Total activity of ornithine aminotransferase in normal blood was 14.1 units and the relative activity of cytosolic and mitochondrial ornithine aminotransferase isozymes were 93.6% and 2.4%, respectively. 3. The reticulocyte count in phenylhydrazine-treated rabbit increased to about 35 times on day 5. Total ornithine aminotransferase activity way maximal on day 5, about 123.4 units which is about 9.4 fold higher than normal value. The relative activity of cytosolic and mitochondrial ornithine aminotransferase isozymes were 97.3% and 2.7%. respectively. 4. The increase in total ornithine aminotransferase activity during induction of reticulocytosis was due primarily to the 9.1 fold increase in the activity cytosolic isozyrrie(Ks=1.1530; Kd=0.0096 ; t½=72.2 hours). 5. The activity of the mitochondrial isozyme increased to about 22.1 times its normal value (Ks=O.0339 ; Kd=0.0102 ; t½=67.9 hours). 6. The rate constants following recovery from phenylhydrazine treatment were; Ks=0.43 82; Kd=0.0332 ; t½=20.9 hours for mitochondrial isozyme.

遺傳工學的 利用을 위한 人體細胞培養에 대한 硏究(Ⅰ) : 人體包皮細胞培養에 影響을 주는 因子에 대하여 Human Foreskin CellCulture

高光三,金盛植,車鍾希,李根培,金成原 조선의대 1983 The Medical Journal of Chosun University Vol.8 No.1