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Electrophoretic Heterogeneity of Portunus trituberculatus Hemocyanin Subunits
유병선,김상복,이강석,이정호,양규환,Yoo, Byung-Sun,Kim, Sang-Bog,Lee, Kang-Suk,Lee, Jeong-Ho,Yang, Kyu-Hwan 생화학분자생물학회 1989 한국생화학회지 Vol.22 No.1
본 연구에서는 꽃게 헤모시아닌 monomer의 전기영동적 이질성에 관해서 여러가지 전기영동 방법을 이용하여 연구하였다. 꽃게 헤모시아닌은 SDS를 포함한 전기영동에서 75,000 정도의 분자량을 갖는 단일 밴드를 나타냈다. 따라서 꽃게 헤모시아닌 구성체는 분자량 크기에 따른 이질성은 보이지 않았다. Discontinuous-polyacrylamide gel electrophoresis (PAGE) 방법에 의한 전기영동 결과 EDTA가 첨가되지 않은 5% 젤에서는 단일 monomer 밴드와 polymer 밴드들이 나타났다. 반면에 1 mM EDTA를 첨가한 discontinuous-PAGE에서는 monomer로 보이는 이중 밴드를 나타내어 꽃게 헤모시아닌이 이질적인 성분으로 구성되어 있을 가능성을 보였다. 한편 acrylamide 농도를 높게 하여 설시한 discontinuous-PAGE의 경우 네개의 monomer 밴드를 나타냈다. SDS-PAGE에서 분자량 차이에 의한 monomer의 이질성은 보이지 않았지만 조건을 달리한 discontinuous-PAGE에서 꽃게 헤모시아닌의 monomer가 최소한 네종류인 것이 밝혀졌다. 이러한 사실은 continuous-PAGE에서 확실히 나타났다. Dissociating buffer에서 실시한 continuous-PAGE에서도 네개의 monomer 밴드가 나타났는데 이를 각각 전기영동도에 따라 subunit I, II, III, IV라 명명하였다. Subunit heterogeneity of Portunus trituberculatus hemocyanin was investigated using various electrophoretic systems.SDS polyacrylamide gel electrophoresis (PAGE) was performed to investigate size homogeneity of hemocyanin. The electrophoretic pattern of purified hemocyanin with detergent, SDS in 7.5% gel revealed a single band with a molecular weight of about 75,000. The purified hemocyanin was then subjected to alkaline electrophoresis without detergent. Discontinuous-PAGE of hemocyanin in 5% gel showed single monomer band and higher polymeric bands. Discontinuous-PAGE containing 1 mM EDTA in gel and sample, however, showed two monomeric bands, indicating that the component of hemocyanin might be heterogeneous one. Discontinuous-PAGE in 7.5% acrylamide concentration showed at least four monomeric bands. Inorder to confirm the electrophoretic heterogeneity of Portunus trituberculatus hemocyanin subunits, continuous-PAGE was performed. Continuous-PAGE in a dissociating buffer(Tris/HCl-EDTA buffer pH 8.9)showed that monomeric subunits of Portunus trituberculatus hemocyanin were resolved into four distinct bands, designated as I, II, III, and IV, which agreed with the results of Discontinuous-PAGE in 7.5% gel.
꽃게 헤모시아닌 구성체의 전기영동적 이질성에 관한 연구
유병선,김상복,이강석,이정호,양규환 ( Byung Sun Yoo,Sang Bob Kim,Kang Suk Lee,Jeong Ho Lee,Kyu Hwan Yang ) 생화학분자생물학회 1989 BMB Reports Vol.22 No.1
Subunit heterogeneity of Portunus trituberculatus hemocyanin was investigated using various electrophoretic systems. SDS polyacrylamide gel electrophoresis (PAGE) was performed to investigate size homogeneity of hemocyanin. The electrophoretic pattern of purified hemocyanin with detergent, SDS in 7.5% gel revealed a single band with a molecular weight of about 75,000. The purified hemocyanin was then subjected to alkaline electrophoresis without detergent. Discontinuous-PAGE of hemocyanin in 5% gel showed single monomer band and higher polymeric bands. Discontinuous-PAGE containing 1 mM EDTA in gel and sample, however, showed two monomeric bands, indicating that the component of hemocyanin might be heterogeneous one. Discontinuous-PAGE in 7.5% acrylamide concentration showed at least four monomeric bands. In order to confirm the electrophoretic heterogeneity of Portunus trituberculatus hemocyanin subunits, continuous-PAGE was performed. Continuous-PAGE in a dissociating buffer (Tris/HCI-EDTA buffer pH 8.9) showed that monomeric subunits of Portunus trituberculatus hemocyanin were resolved into four distinct bands, designated as I, II, III, and IV, which agreed with the results of Discontinuous-PAGE in 7.5% gel.