Platform BioPark Archamps, Archamps, France.
Institute for Advanced Biosciences, CR Inserm U1209, CNRS UMR 5309, University of Grenoble-Alpes, Grenoble, France.
Proteomics. 2018 Feb;18(3-4). doi: 10.1002/pmic.201700368. Epub 2018 Feb 2.
The cuticle is a biological composite material consisting principally of N-acetylglucosamine polymer embedded in cuticular proteins (CPs). CPs have been studied and characterized by mass spectrometry in several cuticular structures and in many arthropods. Such analyses were carried out by protein extraction using SDS followed by electrophoresis, allowing detection and identification of numerous CPs. To build a repertoire of cuticular structures from Bombyx mori, Apis mellifera and Anopheles gambiae the use of SDS and electrophoresis was avoided. Using the combination of hexafluoroisopropanol and of a surfactant compatible with MS, a high number of CPs was identified in An. gambiae wings, legs and antennae, and in the thoracic integument cuticle of Ap. mellifera pupae. The exoskeleton analysis of B. mori larvae allowed to identify 85 CPs from a single larva. Finally, the novel proteomics approach was tested on cuticles left behind after the molt from the fourth instar of Acyrthosiphon pisum. Analysis of these cast cuticles allowed to identify 100 Ac. pisum CPs as authentic cuticle constituents. These correspond to 68% of the total putative CPs previously annotated for this pea aphid. While this paper analyzes only the recovered cuticular proteins, peptides from many other proteins were also detected.
表皮是一种生物复合材料，主要由嵌入表皮蛋白 (CPs) 中的 N-乙酰葡萄糖胺聚合物组成。已经通过质谱分析研究和表征了几种表皮结构和许多节肢动物中的 CPs。这种分析是通过 SDS 进行蛋白质提取然后进行电泳来进行的，允许检测和鉴定许多 CPs。为了从家蚕、蜜蜂和冈比亚按蚊构建表皮结构的目录，避免使用 SDS 和电泳。通过使用六氟异丙醇和与 MS 兼容的表面活性剂的组合，在冈比亚按蚊的翅膀、腿和触角以及蜜蜂蛹的胸部体壁表皮中鉴定出大量的 CPs。家蚕幼虫的外骨骼分析允许从单个幼虫中鉴定出 85 种 CPs。最后，该新的蛋白质组学方法在豌豆蚜第四龄蜕皮后留下的蜕皮上进行了测试。对这些蜕下的表皮进行分析，鉴定出 100 种豌豆蚜 CPs 为真正的表皮成分。这些对应于之前为这种豌豆蚜注释的总假定 CPs 的 68%。虽然本文仅分析了回收的表皮蛋白，但也检测到了许多其他蛋白质的肽。
