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非绿色质体包膜膜的制备与表征

Preparation and characterization of envelope membranes from nongreen plastids.

作者信息

Alban C, Joyard J, Douce R

机构信息

Laboratoire de Physiologie Cellulaire Végétale, U. A. Centre National de la Recherche Scientifique 576, 85X, F-38041 Grenoble-Cédex, France.

出版信息

Plant Physiol. 1988 Nov;88(3):709-17. doi: 10.1104/pp.88.3.709.

DOI:10.1104/pp.88.3.709
PMID:16666372
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1055649/
Abstract

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.

摘要

我们已经开发出一种可靠的方法，用于从菜花（甘蓝）芽质体和梧桐（假挪威槭）细胞造粉体中纯化包膜膜。在纯化的完整质体被破坏后，通过在线性蔗糖梯度上离心来实现包膜膜的分离。通过单半乳糖基二酰基甘油合酶的存在，鉴定出一种密度为每立方厘米1.122克且含有类胡萝卜素的膜组分作为质体包膜。使用针对菠菜叶绿体包膜多肽E24和E30产生的抗体，我们证明了该包膜组分中同时存在外膜和内膜。从梧桐和菜花质体的包膜组分中鉴定出的主要多肽在免疫学上为磷酸转运体。在菜花和梧桐质体的包膜膜中，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。从菜花芽质体和梧桐造粉体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的酶、酰基辅酶A硫酯酶和酰基辅酶A合成酶。这些结果表明，非绿色质体的包膜膜与叶绿体包膜膜具有高度同源性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c658/1055649/a227c025368d/plntphys00633-0214-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c658/1055649/474199c9e2a1/plntphys00633-0213-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c658/1055649/74cd631fa5b6/plntphys00633-0213-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c658/1055649/e2f2fb50038b/plntphys00633-0214-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c658/1055649/a227c025368d/plntphys00633-0214-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c658/1055649/474199c9e2a1/plntphys00633-0213-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c658/1055649/74cd631fa5b6/plntphys00633-0213-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c658/1055649/e2f2fb50038b/plntphys00633-0214-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c658/1055649/a227c025368d/plntphys00633-0214-b.jpg

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Enzymic capacities of purified cauliflower bud plastids for lipid synthesis and carbohydrate metabolism.纯化花椰菜芽质体中脂质合成和碳水化合物代谢的酶活性。
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