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        대장균 Strain B 와 B / r 에 있어서 Acyl Phosphatidyl Glycerol 의 생합성 기작에 관한 연구

        조기승,홍승덕,장정순,이강석 ( Key Seung Cho,Seung Duk Hong,Chung Soon Chang,Kang Suk Lee ) 생화학분자생물학회 1975 BMB Reports Vol.8 No.1

        The present study has shown that phosphatidyl glycerol was converted to 3-sn-phosphatidyl-1`-(3`-acyl)-sn-glycerol(acyl phosphatidyl glycerol) by cell free extracts of Escherichia coli B and B/r. In this process, phosphatidyl glycerol was esterified directly rather than degraded to simpler compounds which are subsequently incorporated into acyl phosphatidyl glycerol. The enzyme, $quot;acyl phosphatidyl glycerol synthetase$quot;, which catalyzes the conversion of phosphatidyl glycerol to acyl phosphatidyl glycerol was found to reside solely in the praticulate fraction of E. coli homogenates. Two separate reactions were identified where one molecule of acyl phosphatidyl glycerol was synthesized from two molecules of phosphatidyl glycerol, in which phosphatidyl glycerol acted both as an acyl acceptor and a donor. In the other reaction, phosphatidyl ethanolamine appears to act as an acyl donor to synthesize acyl phosphatidyl glycerol. The formation of acyl phosphatidyl glycerol has an optimum pH value at pH 7.0 and calcium concentration of 6.0 mM. These results show a new pathway for the turnover of phosphatidyl glycerol in Escherichia coli.

      • SCIESCOPUSKCI등재

        적혈구 막에 관한 연구 ( 1 ) Palmityoyl Carnitine 에 의한 소 적혈구 막의 용해

        이강순,장정순,조기승,이강석 ( Kang Soon Rhee,Chung Soon Chang,Key Seung Cho,Kang Suk Lee ) 생화학분자생물학회 1975 BMB Reports Vol.8 No.1

        The mechanism of solubilization of the ox-erythrocyte membrane was investigated with palmitoyl carnitine. When the membrane was treated with palmitoyl carnitine, the membrane protein, phospholipids and cholesterol were completely solubilized. It was confirmed that lipids were not liberated from membrane lipoprotein and not form micelles with palmitoyl carnitine. As a consequence, membrane was solubilized as a form of lipoprotein. In the present study, the results indicated that the erythrocyte membrane was solubilized by involvement of ionic and hydrophobic interaction of membrane lipoprotein with palmitoyl carnitine. The solubility of erythrocyte membrane depended upon the degree of the hydrophobic binding with palmitoyl carnitine. In the complete solubilized membrane, the activities of ATPase (Na^+, K^+-dependent) and glucose-6-phosphatase were not affected or rather slightly increased of 28% and 56% as compared with intact membrane, respectively. The effect of pH on solubilized membrane protein showed that 92% of protein was precipitated at pH 5.0. In polyacrylamide gel electrophoresis with Tris-glycine buffer, pH 8.3, solubilized membrane protein was separated into 8 bands with molecular weight ranged from 450,000 to 70,000.

      • The Mechanism of Acyl Phosphatidyl Glycerol Biosynthesis in Escherichia coli B and B/r

        조기승,홍승덕,장정순,이강석,Cho, Key-Seung,Hong, Seung-Duk,Chang, Chung-Soon,Lee, Kang-Suk 생화학분자생물학회 1975 한국생화학회지 Vol.8 No.1

        본 연구는 대장균 strain B와 B/r(Escherichia coli Band B/r)의 세포막 분획에의 해서 phosphatidyl glycerol이 3-sn-phosphatidyl-1'-(3'-acyl(-sn-glycerol(acyl phosphatidyl glycerol 로 전환되는 기작을 밝혔다. 이 과정에서 phosphatidyl glycerol은 더 간단한 화합물로 분해된 후 acyl phosphatidyl glycerol로 합성되는 것이 아니라 직접 acyl group이 옮겨져서 ester 결합을 하는 것으로 생각된다. Phosphatidyl glycerol이 acyl phosphatidyl glycerol로 전환되는 반응을 촉매하는 효소인 acyl phosphatidyl glycerol synthetase는 E. coli homogenate의 세포막 분획에 존재하는 것으로 밝혀졌다. Phosphatidyl glycerol이 acyl phosphatidyl glycerol로 전환됨에 있어서 두 개의 다른 반응 기작을 밝혔다. 두 분자의 phosphatidyl glycerol이 한 분자의 acyl phosphatidyl glycerol 을 합성하는 즉 phosphatidyl glycerol이 acyl group의 공여체와 수용체로 각각 작용하는 반응과 다른 반응은 phosphatidyl glycer이 이 수용체로 phosphatidyl ethanolamine이 acyl group 공여체로 작용하는 기작이다. Acyl phosphatidyl glycerol 생합성에 있어서 최적 pH 는 7.0이며 최적 calcium ion 농도는 6.0mM 이었다. 이 실험 결과는 대장균의 phosphatidyl glycerol의 새로운 대사과정을 밝혔다. The present study has shown that phosphatidyl glycerol was converted to 3-sn-phosphatidyl-1'-(3'-acyl)-sn-glycerol (acyl phosphatidyl glycerol) by cell free extracts of Escherichia coli Band B/r. In this process, phosphatidyl glycerol was esterified directly rather than degraded to simpler compounds which are subsequently incorporated into acyl phosphatidyl glycerol. The enzyme, "acyl phosphatidyl glycerol synthetase", which catalyzes the conversion of phosphatidyl glycerol to acyl phosphatidyl glycerol was found to reside solely in the praticulate fraction of E. coli homogenates. Two separate reactions were identified where one molecule of acyl phosphatidyl glycerol was synthesized from two molecules of phosphatidyl glycerol, in which phosphatidyl glycerol acted both as an acyl acceptor and a donor. In the other reaction, phosphatidyl ethanolamine appears to act as an acyl donor to synthesize acyl phosphatidyl glycerol. The formation of acyl phosphatidyl glycerol has an optimum pH value at pH 7.0 and calcium concentration of 6.0 mM. These results show a new pathway for the turnover of phosphatidyl glycerol in Escherichia coli.

      • 적혈구 막에 관한 연구(I) Palmitoyl Carnitine에 의한 소 적혈구 막의 용해

        이강순,장정순,조기승,이강석,Rhee, Kang-Soon,Chang, Chung-Soon,Cho, Key-Seung,Lee, Kang-Suk 생화학분자생물학회 1975 한국생화학회지 Vol.8 No.1

        소의 적혈구막을 palmitoyl carnitine으로 처리하였을 때 단백질, 인지질 및 cholesterol이 동시에 완전히 용해되었고 palmitoyl carnitine에의 한 지질의 유리 및 micelle의 형성이 없는 것으로 보아 적혈구막은 lipoprotein 형태로 용해됨을 알았다. 소 적혈구막의 용해는 palmitoyl carnitine이 막의 lipoprotein과의 친수성 결합과 소수성 결합을 하므로써 용해되며 용해도는 소수성 결합 정도에의 존한다고 본다. Palmitoyl carnitine에의 한 혈구막 용해에 있어서 ATPase($Na^+$, $K^+$ dependent)와 glucose-6-phosphatase의 활성도는 저해 받지 않았으며 오히려 palmitoyl carnitine의 농도에 따라 각각 28-43%, 56-110%가 증가하였다. 용해된 막단백질에 있어 pH에 따른 용해도의 상태는 pH 5.0에서 92%의 단백질이 침전되었다. 용해된 막단백질을 Tris-glycine buffer, pH 8.3로 polyacrylamide gel electrophoresis를 행하였을 때 분자량이 450,000-70,000인 8개의 band로 분리되었다. The mechanism of solubilization of the ox-erythrocyte membrane was investigated with palmitoyl carnitine. When the membrane was treated with palmitoyl carnitine, the membrane protein, phospholipids and cholesterol were completely solubilized. It was confirmed that lipids were not liberated from membrane lipoprotein and not form micelles with palmitoyl carnitine. As a consequence, membrane was solubilized as a form of lipoprotein. In the present study, the results indicated that the erythrocyte membrane was solubilized by involvement of ionic and hydrophobic interaction of membrane lipoprotein with palmitoyl carnitine. The solubility of erythrocyte membrane depended upon the degree of the hydrophobic binding with palmitoyl carnitine. In the complete solubilized membrane, the activities of ATPase($Na^+$, $K^+$-dependent) and glucose-6-phosphatase were not affected or rather slightly increased of 28% and 56% as compared with intact membrane, respectively. The effect of pH on solubilized membrane protein showed that 92% of protein was precipitated at pH 5.0. In polyacrylamide gel electrophoresis with Tris-glycine buffer, pH 8.3, solubilized membrane protein was separated into 8 bands with molecular weight ranged from 450, 000 to 70, 000.

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