Trnka M, Peschek G A
Biochem Biophys Res Commun. 1986 Apr 14;136(1):235-41. doi: 10.1016/0006-291x(86)90899-5.
Membranes were isolated from the cyanobacterium Anacystis nidulans by French press extrusion of lysozyme-treated cells. The membranes were solubilized with sodium dodecylsulfate and subjected to denaturing polyacrylamide gel electrophoresis. Separated polypeptides were transferred to nitrocellulose by Western blotting, and incubated with antibodies against aa3-type cytochrome oxidase of Paracoccus denitrificans; antibodies against subunits I and II, and against the holoenzyme, were used and gave pronounced complementary cross reaction with two of the Anacystis membrane polypeptides corresponding to molecular weights of approximately 55,000 and 32,000, respectively. From this we conclude that an aa3-type cytochrome oxidase is present in Anacystis nidulans as was previously suggested from spectral evidence (G.A.Peschek, Biochim.Biophys.Acta 635 (1981) 470-475), and that this enzyme is composed of at least two subunits with apparent homology to subunits I and II of the corresponding Paracoccus cytochrome oxidase.
通过对经溶菌酶处理的细胞进行法国压榨挤出，从蓝藻集胞藻中分离出膜。用十二烷基硫酸钠使膜溶解，并进行变性聚丙烯酰胺凝胶电泳。通过蛋白质印迹法将分离的多肽转移到硝酸纤维素膜上，并用抗反硝化副球菌aa3型细胞色素氧化酶的抗体进行孵育；使用了针对亚基I和II以及全酶的抗体，它们与集胞藻膜上两条分子量分别约为55,000和32,000 的多肽产生明显的互补交叉反应。由此我们得出结论，正如之前光谱证据所表明的那样（G.A.佩施克，《生物化学与生物物理学报》635 (1981) 470 - 475），集胞藻中存在aa3型细胞色素氧化酶，并且这种酶至少由两个亚基组成，这些亚基与相应反硝化副球菌细胞色素氧化酶的亚基I和II具有明显的同源性。
