Proteomic and biochemical evidence support a role for transport vesicles and endosomes in stress response and secondary metabolism in Aspergillus parasiticus.
Proteomic and biochemical evidence support a role for transport vesicles and endosomes in stress response and secondary metabolism in Aspergillus parasiticus.
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DOI：
10.1021/pr2006389
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发表时间：
2012-02-03
影响因子：
4.4
通讯作者：
Roze, Ludmila V.
中科院分区：
文献类型：
--
作者：
Linz, John E.;Chanda, Anindya;Hong, Sung-Yong;Whitten, Douglas A.;Wilkerson, Curtis;Roze, Ludmila V.
黄曲霉毒素是已知的最强的天然致癌物之一。先前的研究表明，在丝状真菌寄生曲霉中的内体携带催化黄曲霉毒素合成的最后两个步骤的酶，并且这些结构也在黄曲霉毒素储存和输出中发挥作用。我们假设核内体包含一个完整的功能性黄曲霉毒素生物合成途径。为了证实这一假设，我们从A.寄生菌在黄曲霉毒素诱导和非诱导条件下生长。我们还将黄曲霉毒素途径中间体添加到V级分中以测试黄曲霉毒素途径酶的功能状态。V级分中蛋白质的高通量LC/MS/MS分析检测到8种高可靠性的黄曲霉毒素酶和8种较低可靠性的其他酶，表明存在大多数黄曲霉毒素途径酶。纯化的V组分合成黄曲霉毒素，并且添加途径中间体versicolorin A增加黄曲霉毒素合成，证实V组分中的中晚期黄曲霉毒素酶是功能性的。特别重要的是，蛋白质组学和生物化学分析强烈表明，额外的次级代谢途径以及参与响应热，渗透压和氧化应激的蛋白质被安置在V级分。
Aflatoxin is among the most potent naturally occurring carcinogens known. Previous studies demonstrated that endosomes in the filamentous fungus Aspergillus parasiticus carry enzymes that catalyze the final two steps in aflatoxin synthesis and these structures also play a role in aflatoxin storage and export. We hypothesized that endosomes house a complete and functional aflatoxin biosynthetic pathway. To address this hypothesis, we purified a cellular fraction containing endosomes, transport vesicles, and vacuoles (V fraction) from A. parasiticus grown under aflatoxin inducing and non-inducing conditions. We also added (fed) aflatoxin pathway intermediates to V fraction to test the functional status of aflatoxin pathway enzymes. High throughput LC/MS/MS analysis of proteins in V fraction detected 8 aflatoxin enzymes with high reliability and 8 additional enzymes at lower reliability suggesting that most aflatoxin pathway enzymes are present. Purified V fraction synthesized aflatoxin and addition of the pathway intermediate versicolorin A increased aflatoxin synthesis confirming that middle and late aflatoxin enzymes in V fraction are functional. Of particular significance, proteomic and biochemical analysis strongly suggested that additional secondary metabolic pathways as well as proteins involved in response to heat, osmotic, and oxidative stress are housed in V fraction.
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影响因子：
3.4
作者：
Duclos, Sophie;Clavarino, Giovanna;Desjardins, Michel
通讯作者：
Desjardins, Michel
影响因子：
--
作者：
Hong, Sung-Yong;Linz, John E.
通讯作者：
Linz, John E.
DOI：
10.1016/j.bbrc.2005.12.077
发表时间：
2006-02-17
影响因子：
3.1
作者：
Higuchi, Y;Nakahama, T;Kitamoto, K
通讯作者：
Kitamoto, K
影响因子：
4.8
作者：
Roze, LV;Miller, MJ;Linz, JE
通讯作者：
Linz, JE
影响因子：
2.2
作者：
Chanda, Anindya;Roze, Ludmila V.;Pastor, Alicia;Frame, Melinda K.;Linz, John E.
通讯作者：
Linz, John E.