Proteomic analysis of lipid body from the alkenone-producing marine haptophyte alga Tisochrysis lutea.
Proteomic analysis of lipid body from the alkenone-producing marine haptophyte alga Tisochrysis lutea.
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DOI：
10.1002/pmic.201500010
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发表时间：
2015-12
期刊：
影响因子：
3.4
通讯作者：
Shiraiwa, Yoshihiro
中科院分区：
文献类型：
--
作者：
Shi, Qing;Araie, Hiroya;Bakku, Ranjith Kumar;Fukao, Yoichiro;Rakwal, Randeep;Suzuki, Iwane;Shiraiwa, Yoshihiro
脂体是许多生物体中公认的细胞碳和能量储存细胞器。在海洋触生藻类Tsochrysis lutea中观察到Lbs，这种藻能产生特殊的脂类，如具有2-4个反式双键的长链(C37-C40)酮(烯酮)。在本研究中，我们成功地开发了一种从黄曲霉中分离Lb的改良方法。分离的LBS没有叶绿素自动荧光，也没有被最丰富的细胞蛋白核酮糖-1，5-二磷酸羧化酶/加氧酶污染，从而证实了LBS的纯度。通过GC-MS分析发现，由于烷基化合物主要以烷基化合物为主，因此将其命名为“烷基化合物(AB)”更为合适。提取的AB蛋白用1DE(SDS-PAGE)和MS/MS相结合的方法进行蛋白质鉴定，并在国家生物技术信息中心使用BLAST工具进行注释。最多可鉴定出514个蛋白质。同源性搜索确定了三个主要的蛋白质，V-ATPase，一个在其他产生烯酮的触须中发现的假想蛋白质EMIHUDRAFT_465517，以及一个与脂筏相关的SPFH结构域的蛋白质。我们的数据表明，黑斑潜蝇的AB被一层来自内质网或内质网衍生的四层膜叶绿体的脂膜所包围，并作为烯烃和烯烃的储存场所。
Lipid body (LB) is recognized as the cellular carbon and energy storage organelle in many organisms. LBs have been observed in the marine haptophyte alga Tisochrysis lutea that produces special lipids such as long‐chain (C37‐C40) ketones (alkenones) with 2–4 trans‐type double bonds. In this study, we succeeded in developing a modified method to isolate LB from T. lutea. Purity of isolated LBs was confirmed by the absence of chlorophyll auto‐fluorescence and no contamination of the most abundant cellular protein ribulose‐1,5‐bisphosphate carboxylase/oxygenase. As alkenones predominated in the LB by GC‐MS analysis, the LB can be more appropriately named as “alkenone body (AB).” Extracted AB‐containing proteins were analyzed by the combination of 1DE (SDS‐PAGE) and MS/MS for confident protein identification and annotated using BLAST tools at National Center for Biotechnology Information. Totally 514 proteins were identified at the maximum. The homology search identified three major proteins, V‐ATPase, a hypothetical protein EMIHUDRAFT_465517 found in other alkenone‐producing haptophytes, and a lipid raft‐associated SPFH domain‐containing protein. Our data suggest that AB of T. lutera is surrounded by a lipid membrane originating from either the ER or the ER‐derived four layer‐envelopes chloroplast and function as the storage site of alkenones and alkenes.
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