간질(Fasciola hepatica)의 Aspartate 및 Alanine Aminotransferase에 관하여

박선효(Sun Hyo Park),권년수(Nyon Soo Kwon),이희성(Hi Sung Lee),송철용(Chul Yong Song) 대한기생충학열대의학회 1983 The Korean Journal of Parasitology Vol.21 No.1

Phenylhydrazine으로 유발된 빈혈에 대한 방어기전으로서의 Superoxide Dismutase의 역할

이광재,권년수,이동욱,이희성 중앙대학교 의과대학 의과학연구소 1983 中央醫大誌 Vol.8 No.3

Cytotoxic superoxide radical(O_2^-) is a common intermediate of oxygen reduction in the respiring cells and therefore these cells should be protected against the cytotoxic effects of superoxide radicals. Superoxide dismutase(EC 1.15.1.1) is an enzyme that makes a defence against the toxicity of superoxide radical. At present time, three types of superoxide dismutase(SOD), copper and zinc-containing, manganese containing and iron containing enzyme, have been isolated from both eukaryotic and prokaryotic cells. In this study, we have carried out an experiment to establish the general patterns of cynthesis and degradation of SOD in rabbit that was induced hemolytic anemia by phenylhydrazine and normal, sequential measurements of activity of SOD in the cytosolic and mitochondrial fraction of normal and phenylhydrazine treated rabbits were conducted. The kinetic study of rabbit red blood cell(RBC) during cytodifferentiation provides a unique system which permitted a comparison of the isozymes' turnover rates in vivo. Also, in order to observe the role of SOD as protecting mechanism against O_2^- radicals in this experimental system, these data were treated according to kinetic method of the Kopelovich. The activity of SOD was measured by the method of McCord and Fridovich. The experimental results are summarized as follows; 1. The activity of cytosolic and mitochondrial SOD of normal rabbit RBC was 42 units and 1.5 units per ml RBC, respectively. 2. The reticulocyte counts of normal rabbit RBC was approximately 2.5% and administration of phenylhydrazine resulted in rapid and profound reticulocytosis, i.e., about 63.5% on day 6 and essentially 100% on day 7. 3. The hematocrit of normal rabbit blood was 36%, but after treatment of phenylhydrazine, it was decreased to 20.2% in the value on 4th day. 4. In the phenylhydrazine treated rabbit RBC, activity of cytosolic SOD was maximal on day 5, about 108 units per ml RBC which is about 2.6 fold higheh than the normal value. At induction period, rate constants of synthesis(K_s) and degradation(K_d) were 3.9312 and 0.0364, respectively, the half life was 19.04hrs. K_s and K_d following recovery from phenylhydrazine treatment were 0.5781 and 0.0141, respectively, and the half life was 49.0hrs. 5. The activity of mitochondrial SOD was maximal on day 5, about 5.5 fold higher than the normal value. During induction period, rate constants of synthesis and degradation were 0.1640 and 0.0200, respectively, and the half lief was 34.65 hrs K_s and K_d following recovery from phenylhydrazine treatment were 0.0384 and 0.0256, respectively, and the half life was 27.1 hrs.

토끼 뇌조직의 Superoxide Dismutase의 정제 및 성상

최문식,권년수,이희성 중앙대학교 의과대학 의과학연구소 1982 中央醫大誌 Vol.7 No.4

Distribution and some properities of superoxide dismutase was investigated in rabbit brain. Rabbit brain was fractionated by differential centrifugation into mitochondrial and cytosolic fractions. Cytosolic superoxide dismutase was purified by ammonium sulface precioitation, treatment with ethanol-chloroform mixture, and DEAE-cellulose column chromatography. The activity of superoxide dismutase was measured by method of McCord and Fridovich. The results are summarized as follows. 1. Rabbit brain contains two types of superoxide dismutase, one in the mitochondrial and the other in the cytosol. The mitochondrial superoxidase was inactivated by treatment with mixture of ethanol-chloroform whereas the cytosolic superoxide dismutase was not. 2. The activity in homogenate was found to be 93.3 units/g of wet tissue. The distribution of superoxide dismutase in the subcellular organelle showed that 61% of the activity was in cytosolic fraction and 39% in mitochondrial fraction. 3. The molecular weight og cytosolic superoxide dismutase was estimated to be 32,000 by gel filtration. 4. The straviolet absorption spectrum of cytosolic superoxide dismutase indicated a lack of tryptophan. The spectrum of the enzyme in the altraviolet region was similar to the absorption spectrum of phenylalanine and tyrosine. 5. Purified cytosolic superoxide dismutase contained 2 Cu^2+ and 2 Zn^2+ per mole. This enzyme was found to be similar to the other cupro-zine superoxide dismutase which have been isolated from diverse eukaryotes. 6. Mitochondrial superoxide dismutase contained Mn^2+ and was strikingly similar to the superoxide dismutase previously isolated from bacteria. 7. Cyanide at 0.5mM and 5.0 mM inhibited the activity of cytosolic superoxide dismutase 13% amd 93%, repectively. 8. Superoxide dismutase activity in mitochondria was partitioned 38% in the intermenbrane space and 62% in the matrix. 9. The activity of xanthine oxidase in homogenate was fiund to be 333 units/g of wet tissue and the relative of cytosolic and mitochondrial xanthine oxidase was 77% and 23%, respectively.

간질(Fasciola hepatica)의 Krebs Cycle 및 α-glycerophosphate Shuttle에 관한 연구

한인숙,이동욱,권년수,이희성 중앙대학교 의과대학 의과학연구소 1983 中央醫大誌 Vol.8 No.2

This study was carried out to observe the activities and their distributions of several enzymes in Fasciola hepatica. The whole body homogenate was separated into cytosolic, mitochondrial and nucleic fractions by differential centrifugation. The activities of enzymes, NAD-linked α-glycerophosphate dehydrogenase(EC 1.1.1.8), FAD-linked isocitrate dehydrogenase(EC 1.1.1.42), trehalase(EC 3.2.1.28), branched chain amino acid aminotransferase(EC 2.6.1.24) and adenosine deaminase(EC 3.5.4.4) were measured in each fraction. The results were summarized as follows: 1. The activity of NAD-linked α-glycerophosphate dehydrogenase in cytosolic fraction was observed to be 1,998 units per one gram wet tissue, 93% of total activity and its specific activity was 2.1. The activity of FAD-linked α-glycerophosphate dehydrogenase in mitochondrial fraction was founded to be 39 units per one gram of wet tissue. It was fall under 67% of total activity and specific activity was 1.3. The rate of oxidation of α-glycerophosphate dehydrogenase by NAD- and FAD-linked α-glycerophosphate dehydrogenase were remarkable differed. This result indicates that transportation of reducing equivalents into mitochondria from the cytosol does not active. 2. The activity of NAD-linked isocitrate dehydrogenase was not detected in Fasciola hepatica whereas the NADP-linked isocitrate dehydrogenase was existed in both cytosolic and mitochondrial fraction. There activities were comparatively low. Therefore the Krebs cycle seems to be not active in Fasciola hepatica. NADPH produced by the action of NADP-linked isocitrate dehydrogenase may be utilized in the synthesis of fatty acids. 3. Relatively high activity of branched chain amino acid aminotransferase was observed in all fractions, and 81~87% of total activity was localized in cytosolic fraction. The best substrate of the branched chain amino acid aminotransferase among the three amino acids(leucine, valine and isoleucine) was isoleucine, and the worst was valine. 4. The activity of trehalase was not detected, it suggested that trehalose was not utilized by fasciola hepatica. 5. Deamination of adenosine by deaminase was not found.