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Preparation and Characterization of Envelope Membranes from Nongreen Plastids

机译:非绿色塑料质膜的制备与表征

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摘要

We have developed a reliable procedure for the purification of envelope membranes from cauliflower (Brassica oleracea L.) bud plastids and sycamore (Acer pseudoplatanus L.) cell amyloplasts. After disruption of purified intact plastids, separation of envelope membranes was achieved by centrifugation on a linear sucrose gradient. A membrane fraction, having a density of 1.122 grams per cubic centimeter and containing carotenoids, was identified as the plastid envelope by the presence of monogalactosyldiacylglycerol synthase. Using antibodies raised against spinach chloroplast envelope polypeptides E24 and E30, we have demonstrated that both the outer and the inner envelope membranes were present in this envelope fraction. The major polypeptide in the envelope fractions from sycamore and cauliflower plastids was identified immunologically as the phosphate translocator. In the envelope membranes from cauliflower and sycamore plastids, the major glycerolipids were monogalactosyldiacylglycerol, digalactosyldiacylglycerol, and phosphatidylcholine. Purified envelope membranes from cauliflower bud plastids and sycamore amyloplasts also contained a galactolipid:galactolipid galactosyltransferase, enzymes for phosphatidic acid and diacylglycerol biosynthesis, acyl-coenzyme A thioesterase, and acyl-coenzyme A synthetase. These results demonstrate that envelope membranes from nongreen plastids present a high level of homology with chloroplasts envelope membranes.
机译:我们已经开发了一种从花椰菜(Brassica oleracea L.)芽质体和无花果(Acer pseudoplatanus L.)细胞淀粉状体中纯化包膜的可靠方法。破坏纯化的完整质体后,通过在线性蔗糖梯度上离心来分离包膜。通过存在单半乳糖基二酰基甘油合酶,将具有1.122克/立方厘米密度并含有类胡萝卜素的膜级分鉴定为质体包膜。使用针对菠菜叶绿体包膜多肽E24和E30产生的抗体,我们证明了在该包膜部分同时存在外和内包膜。从美国梧桐和花椰菜质体的包膜部分主要多肽被免疫学鉴定为磷酸盐易位。在花椰菜和梧桐质体的包膜中,主要的甘油脂是单半乳糖基二酰基甘油,二半乳糖基二酰基甘油和磷脂酰胆碱。从花椰菜芽质体和梧桐淀粉质中纯化的包膜膜还含有半乳糖脂:半乳糖脂半乳糖基转移酶,磷脂酸和二酰基甘油生物合成酶,酰基辅酶A硫酯酶和酰基辅酶A合成酶。这些结果表明,来自非绿色质体的包膜与叶绿体包膜具有高度的同源性。

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