사람 적혈구에 있어서 극성 대사물질과 막 구성분의 Uptake에 관한 1-Palmitoyllysolecithin과 인삼 Saponin의 영향

정진성,이종호,조기승,주충노,Chung, Jin-Sung,Lee, Jong-Ho,Cho, Key-Seung,Joo, Chung-No 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.2

사람 적혈구에서 [$^{14}C$]-alanine, [$^{14}C$]-glucose, [$^{14}C$]-cholesterol 및 [$^{14}C$]-phosphatidylethanolamine의 흡수를 lysolecithin과 saponin 존재하에서 실험하여 본 결과 다음과 같은 결과를 얻었다. 적혈구를 lysolecithin으로 미리 처리시켰을 때, 상기 물질들의 uptake가 alanine의 경우 최고 111%, glucose는 43%, cholesterol은 48%, phosphatidylethanolamine은 17%의 상승율을 보였고, saponin의 처리에 의해서는 각각 408%, 70%, 28% 및 10%의 상승율을 나타냈다. 두 lytic agent에 의한 상승율은 각각의 최적 농도에서 비교해보면 lysolecithin은 saponin 경우보다 막성분인 cholesterol이나 phosphatidylethanolamine의 uptake에 더 영향을 끼쳤고, 한편 saponin은 alanine과 glucose와 같은 polar metabolite의 uptake에 더 영향을 주는 것으로 확인되었다. The effects of lysolecithin and ginseng saponin on the uptake of [$^{14}C$]-alanine, [$^{14}C$]-glucose, [$^{14}C$]-cholesterol, and [$^{14}C$]-phosphatidylethanolamine to human erythrocytes in the various conditions were as follows. When the human erythrocytes pretreated in the presence of low lytic amount of lysolecithin, the uptake of such polar metabolites and membrane components was enhanced up to [$^{14}C$]-alanine 111%, [$^{14}C$]-glucose 43%, [$^{14}C$]-cholesterol 48%, and [$^{14}C$]-phosphatidylethanolamine 17%, In case of saponin pretreatment, the uptakes of such substances were increased to 408%, 70%, 28%, and 10%, respectively. In comparison with both lytic surfactants under the optimum concentration, lysolecithin was more effective to the uptake of membrane components, cholesterol and phosphatidylethanolamine. On the other hand, saponin was more potent to the polar metabolite uptake into the erythrocytes.

생쥐 간의 Triacylglycerol 생합성에 있어서 Palmitoylcarnitine의 지방산 공여체로서의 역할

이윤경,정진성,조기승,Lee, Youn-Kyung,Chung, Jin-Sung,Cho, Key-Seung 생화학분자생물학회 1986 한국생화학회지 Vol.19 No.3

$^{14}C$-Palmitoylcarnitine을 생쥐 간 균질물과 반응시킨 결과 1시간 후 33%, 3시간 후에 51%, 6시간 후에 최고 61%의 가수분해를 나타냄으로서, 동물체의 간이 acylcarnitine의 중요한 대사부위 임을 나타내 주었다. $^{14}C$-Palmitoylcarnitine ($^{14}C$-pal. carn)에 의한 $^{14}C$-triacylglycerol ($^{14}C$-TG)의 생합성은 0.1 M tris-HCl buffer, pH 7.4 및 반응온도 $20^{\circ}C$에서 각각 최고의 합성율을 나타냈고, ATP, coenzyme A (CoASH), $Ca^{2+}$ 및 $Mg^{2+}$의 cofactor 등은 triacylglycerol 생 합성계에 전혀 영향을 끼치지 않았으며, 이들은 phospholipid 생성계를 활성화시키는 것으로 나타났다. 동일 조건하에서 (U-$^{14}C$)-palmitic acid에 의한 중성지 질의 생합성을 비교하여 본 결과, 반응온도는 $20^{\circ}C$가 적정온도였고, 이 반응계는 ATP, CoASH 및 $Mg^{2+}$의 cofactor를 필요로 했는데 반응시간에 따라 차이를 보여주었다. 즉, 30분 반응에서는 cofactor 존재하에서 $^{14}C$-TG 생성이 증가를 보였으나, 3시간 반응에서는 cofactor가 존재하지 않은 대조구보다 $^{14}C$-phospholipids 생성이 증가함을 나타냈다. 간 균질물의 여러 분획에 있어서 $^{14}C$-pal. carn에 의한 $^{14}C$-TG의 생성을 비교해 본 결과, microsomal cytosol 분획에서 가장 높은 triacylglycerol와 diacylglycerol의 생성율을 보였고, 다음이 mitochondria 분획, nuclei와 cell debris 분획 순이었다. 이상의 결과로 보아 palmitoylcarnitine은 microsome에서 cofactor의 도움 없이 직접 acyl group을 이전시켜 중성지질을 생성하는 것을 확인할 수 있었다. It was confirmed that the important metabolic site of acylcarnitine was liver in mamals from the results of its hydrolysis, in which percent of hydrolysis was 33%, 51 %, and 61% from incubation for 1 h, 3 h, and 6 h, respectively. In the triacylglycerol biosynthesis from (U-$^{14}C$)-palmitolyl-DL-carnitine, the optimum conditions were obtained in 0.1 M tris-HCl buffer, pH 7.4 and $20^{\circ}C$ of incubation temperature. The effect of cofactors, such as $Ca^{2+}$, $Mg^{2+}$, ATP, and coenzyme A reduced the formation of triacylglycerol, rather increased the formation of phospholipid. This result showed that triacylglycerol synthesis from palmitolycarnitine did not need any cofactors. The comparison of the triacylglycerol biosynthesis from (U-$^{14}C$)-palmitic acid with that of (U-$^{14}C$)-palmitoyl-DL-carnitine showed the same optimum conditons in pH and incubation temperature. But in cofactors effect, tracylglycerol synthesis from $^{14}C$-palmitic acid was stimulated in the presence of ATP, CoASH, and $Mg^{2+}$ in 30 min incubation when it was compared with or without of cofactors. When the incubation period was prolonged to 3 h, triacylglycerol synthesis was reduced and, on the other hand, phospholipid synthesis was increased significantly more than 6 times. With the results from the different cell fractions, the highest triacylglycerol biosynthesis was prepresented with microsomal cytosol fraction and next was in order of mitochondrial fraction and nuclei cell debris fraction. As conclusion, firstly, it could be postulated that the direct mobilization of acyl group from palmitoylcarnitine to monoacylglycerol and diacylglycerol resulted triacylglycerol synthesis. Secondly, free fatty acid from hydrolysis of palmitoylcarnitine might from acyl CoA in the presence of ATP and CoASH, and continued the sequencial reactions with diacylglycerol and with Iysophospholipid to form the triacylglycerol and phospholipid, respectively.

사람의 적혈구막의 Palmitoylcarnitine 과 Palmitoyl - Coa Carnitine Palmitine Palmitoyltransferase

조기승,정진성 ( Key Seung Cho,Jin Sung Chung ) 생화학분자생물학회 1982 BMB Reports Vol.15 No.3

An enzyme, palmitoyl-CoA: carnitine palmitoyltransferase (E.C. 2. 3.1.21) was confirmed to catalyze reversively the formation and hydrolysis of ^(14)C-palmitoylcarnitine in human erythrocyte membrane. The optimum pH range of this enzyme was between from pH 7.2 to pH 7.4, and decreased rapidly outside of this pH range. In the several buffer systems tested, the highest enzyme activity was obtained with Tris-HCl buffer. As a cofactor, 5 μmoles of Mg^(2+) ion significantly stimulated the enzyme activity in the experimental conditions. Palmitoylcarnitine formed from this enzyme system or synthesized from other chemical method has activated significantly the membrane bound enzymes, such as adenosine triphosphatase (ATPase) and Glucose-6-Phosphatase (G-6-Pase).

사람의 적혈구막의 Palmitoylcarnitine과 Palmitoyl-CoA : Carnitine Palmitine Palmitoyltransferase의 역할에 관한 연구

조기승,정진성,Cho, Key-Seung,Chung, Jin- Sung 생화학분자생물학회 1982 한국생화학회지 Vol.15 No.3

An enzyme, palmitoyl-CoA: carnitine palmitoyltransferase (E.C. 2.3.1.21) was confirmed to catalyze reversively the formation and hydrolysis of $^{14}C$ -palmitoylcarnitine in human erythrocyte membrane. The optimum pH range of this enzyme was between from pH 7.2 to pH 7.4, and decreased rapidly outside of this pH range. In the several buffer systems tested, the highest enzyme activity was obtained with Tris-HCl buffer. As a cofactor, 5 ${\mu}moles$ of $Mg^{2+}$ ion significantly stimulated the enzyme activity in the experimental conditions. Palmitoylcarnitine formed from this enzyme system or synthesized from other chemical method has activated significantly the membrane bound enzymes, such as adenosine triphosphatase (ATPase) and Glucose-6-Phosphatase (G-6-Pase). 사람의 적혈구막에서 palmitoyl-CoA: carnitine palmitoyltransferase에 의해 $^{14}C$-palmitoylcarnitine이 가역적으로 생성 및 가수분해 됨을 확인하였다. 이 효소의 최적 pH 범위는 pH 7.2에서 pH 7.4 근방으로, 이 pH 범위 외에서는 급속한 효소활성도의 감소를 나타냈다. 몇가지의 완충용액을 시험하여 보았을때 Tris-HCI buffer가 가장 높은 효소활성도를 보였으며, 조요소로서 5 ${\mu}mole$의 $Mg^{2+}$이온이 동일 실험조건하에서 효소의 활성도를 굉장히 증가시켰다. 이 효소계에 의해서나 흑은 화학적인 합성에 의해 만들어진 palmitoyl. carnitine은 세포막에 부착되어 있는 효소인 ATPase나 G-6-Pase와 같은 효소를 상당히 활성화 시켰다.

생쥐 간의 Triacylglycerol 생합성에 있어서 Palmitoylcarnitine 의 지방산 공여체로서의 역활

정진성,조기승,이윤경 생화학분자생물학회 1991 BMB Reports Vol.19 No.3

It was confirmed that the important metabolic site of acylcarnitine was liver in mamals from the results of its hydrolysis, in which percent of hydrolysis was 33%, 51%, and 61% from incubation for 1 h, 3 h, and 6 h, respectively. In the triacylglycerol biosynthesis from (U-^(14)C)-palmitolyl- DL -carnitine, the optimum conditions were obtained in 0.1 M tris-HCl buffer, pH 7.4 and 20℃ of incubation temperature. The effect of cofactors, such as Ca^(2+), Mg^(2+), ATP, and coenzyme A reduced the formation of triacylglycerol, rather increased the formation of phospholipid. This result showed that triacylglycerol synthesis from palmitolycarnitine did not need any cofactors. The comparison of the triacylglycerol biosynthesis from (U-^(14)C)-palmitic acid with that of (U-^(14)C)-palmitoyl-DL-carnitine showed the same optimum conditons in pH and incubation temperature. But in cofactors effect, tracylglycerol synthesis from ^(14)C-palmitic acid was stimulated in the presence of ATP, CoASH, and Mg^(2+) in 30 min incubation when it was compared with or without of cofactors. When the incubation period was prolonged to 3 h, triacylglycerol synthesis was reduced and, on the other hand, phospholipid synthesis was increased significantly more than 6 times. With the results from the different cell fractions, the highest triacylglycerol biosynthesis was prepresented with microsomal cytosol fraction and next was in order of mitochondria) fraction and nuclei cell debris fraction. As conclusion, firstly, it could be postulated that the direct mobilization of acyl group from palmitoylcarnitine to monoacylglycerol and diacylglycerol resulted triacylglycerol synthesis. Secondly, free fatty acid from hydrolysis of palmitoylcarnitine might from acyl CoA in the presence of ATP and CoASH, and continued the sequential reactions with diacylglycerol and with lysophospholipid to form the triacylglycerol and phospholipid, respectively.

사람 적혈구에 있어서 극성 대사물질과 막 구성분의 Uptake 에 관한 1 - Palmitoyllysolecithin 과 인산 Saponin 의 영향

정진성,이종호,조기승,주충노 생화학분자생물학회 1986 BMB Reports Vol.19 No.2

The effects of Lysolecithin and ginseng saponin on the uptake of [^(14)C]-alanine, [^(14)C]-glucose, [^(14)C]-cholesterol, and [^(14)C]-phosphatidylethanolamine to human erythrocytes in the various conditions were as follows. When the human erythrocytes pretreated in the presence of low lytic amount of lysolecithin, the uptake of such polar metabolites and membrane components was enhanced up to [^(14)C]-alanine 111%, [^(14)C]-glucose 43%, [^(14)C]-cholesterol 48%, and [^(14)C]-phosphatidylethanolamine 17%, In case of saponin pretreatment, the uptakes of such substances were increased to 408%, 70%, 28%, and 10%, respectively. In comparison with both lytic surfactants under the optimum concentration, lysolecithin was more effective to the uptake of membrane components, cholesterol and phosphatidylethanolamine. On the other hand, saponin was more potent to the polar metabolite uptake into the erythrocytes.