Proteomic analysis of the acidocalcisome, an organelle conserved from bacteria to human cells.
Proteomic analysis of the acidocalcisome, an organelle conserved from bacteria to human cells.
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从细菌到人类细胞的细胞器保守的酸性蛋白质组学分析。
DOI：
10.1371/journal.ppat.1004555
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发表时间：
2014-12
期刊：
影响因子：
6.7
通讯作者：
Docampo R
中科院分区：
文献类型：
--
作者：
Huang G;Ulrich PN;Storey M;Johnson D;Tischer J;Tovar JA;Moreno SN;Orlando R;Docampo R
酸钙体是存在于从细菌到人类细胞的各种生物体中的酸性细胞器。在这项研究中，从模式生物布氏锥虫中纯化酸钙体，并通过质谱法测定其蛋白质组成。这些结果与我们先前报道的结果沿着表明，酸钙体富含泵和转运蛋白，参与磷酸盐和阳离子的稳态以及钙信号传导。我们验证了七个新的，假定的，酸钙体蛋白质的酸钙体定位（磷酸盐转运蛋白、液泡H+-ATP酶亚基a和d、液泡铁转运蛋白、锌转运蛋白、多胺转运蛋白和酸性磷酸酶），证实了六种先前表征的酸钙体蛋白的存在，并通过表达与其内源基因座中的表位标签融合的五种新蛋白质来验证它们在不同亚细胞区室中的定位，通过免疫荧光显微镜用特异性抗体。通过RNA干扰（RNAi）敲低几种新鉴定的酸钙体蛋白揭示了它们对寄生虫的生存是必不可少的。这些结果提供了一个全面的洞察独特的组成，酸钙体的T。布氏杆菌，一种重要的真核病原体，和直接证据表明，酸钙体是特别适合于聚磷酸盐的积累。酸钙体是从细菌到人类细胞的酸性细胞器，富含多磷酸盐，一种由高能磷酸-酸酐键连接的正磷酸盐单元的聚合物。我们在这里发现，来自布氏锥虫的酸钙体，属于产生非洲昏睡病和长形线虫的生物体，富含参与磷酸盐和阳离子稳态以及钙信号传导的泵、通道和转运蛋白。酸钙体组分和表达的基因与表位标签的蛋白质组学分析验证了一些新的转运蛋白的存在下，和RNA干扰证明了这些细胞器的重要性。
Acidocalcisomes are acidic organelles present in a diverse range of organisms from bacteria to human cells. In this study acidocalcisomes were purified from the model organism Trypanosoma brucei, and their protein composition was determined by mass spectrometry. The results, along with those that we previously reported, show that acidocalcisomes are rich in pumps and transporters, involved in phosphate and cation homeostasis, and calcium signaling. We validated the acidocalcisome localization of seven new, putative, acidocalcisome proteins (phosphate transporter, vacuolar H+-ATPase subunits a and d, vacuolar iron transporter, zinc transporter, polyamine transporter, and acid phosphatase), confirmed the presence of six previously characterized acidocalcisome proteins, and validated the localization of five novel proteins to different subcellular compartments by expressing them fused to epitope tags in their endogenous loci or by immunofluorescence microscopy with specific antibodies. Knockdown of several newly identified acidocalcisome proteins by RNA interference (RNAi) revealed that they are essential for the survival of the parasites. These results provide a comprehensive insight into the unique composition of acidocalcisomes of T. brucei, an important eukaryotic pathogen, and direct evidence that acidocalcisomes are especially adapted for the accumulation of polyphosphate. Acidocalcisomes are acidic organelles conserved from bacteria to human cells that are rich in polyphosphate, a polymer of orthophosphate units linked by high-energy phospho-anyhidride bonds. We found here that acidocalcisomes from Trypanosoma brucei, belonging to the group of organisms that produces African sleeping sickness and nagana, are rich in pumps, channels, and transporters involved in phosphate and cation homeostasis, and calcium signaling. Proteomic analysis of acidocalcisome fractions and expression of genes with epitope tags validated the presence of a number of novel transporters, and RNA interference demonstrated the essentiality of these organelles.
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DOI：
10.1074/mcp.m900038-mcp200
发表时间：
2009-09
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
DeGrasse JA;DuBois KN;Devos D;Siegel TN;Sali A;Field MC;Rout MP;Chait BT
通讯作者：
Chait BT
影响因子：
4
作者：
Adachi, Atsuhiro;Kano, Fumi;Murata, Masayuki
通讯作者：
Murata, Masayuki
Wolf Psort：蛋白质定位预测因子。
影响因子：
14.9
作者：
Horton, Paul;Park, Keun-Joon;Obayashi, Takeshi;Fujita, Naoya;Harada, Hajime;Adams-Collier, C J;Nakai, Kenta
通讯作者：
Nakai, Kenta
影响因子：
1.5
作者：
Chan, Kuan Yoow;Ersfeld, Klaus
通讯作者：
Ersfeld, Klaus
影响因子：
56.9
作者：
Hothorn, Michael;Neumann, Heinz;Mayer, Andreas
通讯作者：
Mayer, Andreas