CISM, University of Firenze, Firenze, Italy.
J Proteome Res. 2010 Apr 5;9(4):1822-33. doi: 10.1021/pr900969k.
Chemical communication in insects is mediated by soluble binding proteins, belonging to two large families, Odorant-binding Proteins (OBPs) and Chemosensory Proteins (CSPs). Recently, evidence has been provided that OBPs are involved in recognition of chemical stimuli. We therefore decided to investigate the expression of OBPs and CSPs in the honeybee at the protein level, using a proteomic approach. Our results are in agreement with previous reports of expression at the RNA level and show that 12 of the 21 OBPs predicted in the genome of the honeybee Apis mellifera and 2 of the 6 CSPs are present in the foragers' antennae, while the larvae express only three OBPs and a single CSP. MALDI mass spectrometry on crude antennal extracts and MALDI profiling on sections of antennae demonstrated that these techniques can be applied to investigate individual differences in the expression of abundant proteins, such as OBPs and CSPs, as well as to detect the presence of proteins in different regions of the antenna. Finally, as part of a project aimed at the characterization of all OBPs and CSPs of the honeybee, we expressed 5 OBPs and 4 CSPs in a bacterial system and measured their affinity to a number of ligands. Clear differences in their binding spectra have been observed between OBPs, as well as CSPs.
昆虫的化学通讯是由可溶性结合蛋白介导的，这些蛋白属于两个大的家族，即气味结合蛋白（OBPs）和化学感受蛋白（CSPs）。最近的证据表明，OBPs 参与了对化学刺激的识别。因此，我们决定采用蛋白质组学方法在蛋白质水平上研究蜜蜂中 OBPs 和 CSPs 的表达。我们的结果与之前在 RNA 水平上的表达报告一致，表明在蜜蜂基因组中预测的 21 个 OBPs 中的 12 个和 6 个 CSPs 中的 2 个存在于觅食者的触角中，而幼虫仅表达 3 个 OBPs 和 1 个 CSP。粗触角提取物的 MALDI 质谱和触角切片的 MALDI 分析表明，这些技术可用于研究丰富蛋白（如 OBPs 和 CSPs）表达的个体差异，以及检测触角不同区域中蛋白的存在。最后，作为旨在表征蜜蜂所有 OBPs 和 CSPs 的项目的一部分，我们在细菌系统中表达了 5 个 OBPs 和 4 个 CSPs，并测量了它们与许多配体的亲和力。我们观察到 OBPs 之间以及 CSPs 之间在结合谱上存在明显差异。
