Olfactory proteins mediating chemical communication in the navel orangeworm moth, Amyelois transitella.
Olfactory proteins mediating chemical communication in the navel orangeworm moth, Amyelois transitella.
复制标题
DOI：
10.1371/journal.pone.0007235
复制
发表时间：
2009-09-30
期刊：
影响因子：
3.7
通讯作者：
Ames JB
中科院分区：
文献类型：
--
作者：
Leal WS;Ishida Y;Pelletier J;Xu W;Rayo J;Xu X;Ames JB
脐橙小食心虫是加州杏树和开心果上最严重的害虫，环境友好的防治方法，如基于信息素的方法，是非常可取的。性信息素的一些成分是不稳定的，可以被副激素取代，副激素可能是基于信息素和信息素检测嗅觉蛋白的分子相互作用而设计的。通过分析嗅觉和非嗅觉组织的提取物，我们鉴定了可能的嗅觉蛋白，通过Edman降解获得了它们的N端氨基酸序列，并用简并引物通过SMART RACE方法克隆了相应的cDNA。此外，我们使用基于已知蛋白质保守序列的简并引物来筛选其他候选嗅觉基因。我们表达了编码一个新鉴定的信息素结合蛋白的基因，通过圆二色谱、荧光和核磁共振进行了分析，并用于结合实验来评估与信息素组分的亲和力。我们从脐橙小虫中克隆了9个嗅觉蛋白的编码基因，包括2个信息素结合蛋白、2个一般气味结合蛋白、1个化学感觉蛋白、1个谷胱甘肽S转移酶、1个触角结合蛋白X、1个感觉神经元膜蛋白和1个气味受体。其中，AtraPBP1在雄性触角中高度丰富。荧光、圆二色谱和核磁共振研究表明，它的构象发生了显著的pH依赖性变化，在中性pH下与信息素组分有很高的亲和力，在低pH下没有结合。
The navel orangeworm, Amyelois transitella Walker (Lepidoptera: Pyralidae), is the most serious insect pest of almonds and pistachios in California for which environmentally friendly alternative methods of control — like pheromone-based approaches — are highly desirable. Some constituents of the sex pheromone are unstable and could be replaced with parapheromones, which may be designed on the basis of molecular interaction of pheromones and pheromone-detecting olfactory proteins. By analyzing extracts from olfactory and non-olfactory tissues, we identified putative olfactory proteins, obtained their N-terminal amino acid sequences by Edman degradation, and used degenerate primers to clone the corresponding cDNAs by SMART RACE. Additionally, we used degenerate primers based on conserved sequences of known proteins to fish out other candidate olfactory genes. We expressed the gene encoding a newly identified pheromone-binding protein, which was analyzed by circular dichroism, fluorescence, and nuclear magnetic resonance, and used in a binding assay to assess affinity to pheromone components. We have cloned nine cDNAs encoding olfactory proteins from the navel orangeworm, including two pheromone-binding proteins, two general odorant-binding proteins, one chemosensory protein, one glutathione S-transferase, one antennal binding protein X, one sensory neuron membrane protein, and one odorant receptor. Of these, AtraPBP1 is highly enriched in male antennae. Fluorescence, CD and NMR studies suggest a dramatic pH-dependent conformational change, with high affinity to pheromone constituents at neutral pH and no binding at low pH.
登录
查看更多内容
DOI：
10.1073/pnas.0501447102
发表时间：
2005-04-12
影响因子：
11.1
作者：
Leal, WS;Chen, AM;Tsuruda, JM
通讯作者：
Tsuruda, JM
影响因子：
5.6
作者：
Damberger, Fred F.;Ishida, Yuko;Wuethrich, Kurt
通讯作者：
Wuethrich, Kurt
DOI：
10.1016/j.bbrc.2005.07.176
发表时间：
2005-10-07
影响因子：
3.1
作者：
Lautenschlager, C;Leal, WS;Clardy, J
通讯作者：
Clardy, J
DOI：
10.1073/pnas.0505340102
发表时间：
2005-09-27
影响因子：
11.1
作者：
Ishida, Y;Leal, WS
通讯作者：
Leal, WS
影响因子：
3.7
作者：
Leal WS;Barbosa RM;Xu W;Ishida Y;Syed Z;Latte N;Chen AM;Morgan TI;Cornel AJ;Furtado A
通讯作者：
Furtado A