Sulfur single-wavelength anomalous diffraction crystal structure of a pheromone-binding protein from the honeybee Apis mellifera L
Sulfur single-wavelength anomalous diffraction crystal structure of a pheromone-binding protein from the honeybee Apis mellifera L
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DOI：
10.1074/jbc.m311212200
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发表时间：
2004-02-06
影响因子：
4.8
通讯作者：
Cambillau, C
中科院分区：
文献类型：
--
作者：
Lartigue, A;Gruez, A;Cambillau, C
信息素结合蛋白（pheromone binding proteins，PBPs）是一种分子量约为13 - 17 kDa的小螺旋蛋白，存在于蛾和其它昆虫的多种感觉器官中。它们参与信息素从感觉淋巴到嗅觉受体的运输。我们在这里报告的晶体结构的PBP（Amel-ASP 1）来源于蜜蜂（意大利蜜蜂）的触角，并在酵母毕赤酵母中表达的重组蛋白。Amel-ASP 1的晶体在pH 5.5下获得使用纳米滴结晶技术与一种新的优化程序，和结构最初解决了与单波长异常衍射技术，使用硫异常分散。Amel-ASP 1的结构已在1.6埃分辨率下得到细化。它的折叠与其他PBP/气味剂结合蛋白的折叠大致相似，呈现六个螺旋和三个二硫键。与家蚕（Sandler，B. H、尼科诺娃湖，Leal，W.美国，和Clardy，J.（2000）Chem.Biol.7，143 - 151）和马德拉白顶菌（Lartigue，A.，Gruez，A.，Spinelli，S.，里维埃，S.，布罗苏特河，Tegoni，M.，和Cambillau，C.（2003）J.Biol.Chem.278，30213 - 30218），延伸的C末端折叠到蛋白质中并形成内部疏水腔的壁。它的主链基团与一个偶然的配体，正丁基苯磺酰胺（一种用于塑料的添加剂）建立了两个氢键。然而，这种结合模式可能模仿信息素共混物组分之一所使用的结合模式，并说明了PBPs的结合多功能性。
Pheromone binding proteins (PBPs) are small helical proteins (similar to13-17 kDa) present in several sensory organs from moth and other insect species. They are involved in the transport of pheromones from the sensillar lymph to the olfactory receptors. We report here the crystal structure of a PBP (Amel-ASP1) originating from the honeybee (Apis mellifera) antennae and expressed as recombinant protein in the yeast Pichia pastoris. Crystals of Amel-ASP1 were obtained at pH 5.5 using the nanodrops technique of crystallization with a novel optimization procedure, and the structure was solved initially with the single-wavelength anomalous diffraction technique using sulfur anomalous dispersion. The structure of Amel-ASP1 has been refined at 1.6-Angstrom resolution. Its fold is roughly similar to that of other PBP/odorant binding proteins, presenting six helices and three disulfide bridges. Contrary to the PBPs from Bombyx mori (Sandler, B. H., Nikonova, L., Leal, W. S., and Clardy, J. (2000) Chem. Biol. 7, 143-151) and Leucophea maderae (Lartigue, A., Gruez, A., Spinelli, S., Riviere, S., Brossut, R., Tegoni, M., and Cambillau, C. (2003) J. Biol. Chem. 278,30213-30218), the extended C terminus folds into the protein and forms a wall of the internal hydrophobic cavity. Its backbone groups establish two hydrogen bonds with a serendipitous ligand, n-butyl-benzene-sulfonamide, an additive used in plastics. This mode of binding might, however, mimic that used by one of the pheromonal blend components and illustrates the binding versatility of PBPs.