Gomes Laidson Paes, Gouveia E Silva Catarina, Gaillard Jean-Charles, Armengaud Jean, Coelho Ana Varela
Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Oeiras, Portugal.
Département Médicaments et Technologies pour la Santé, Université Paris-Saclay, Bagnols-sur-Cèze, France.
Methods Mol Biol. 2022;2450:583-597. doi: 10.1007/978-1-0716-2172-1_31.
Proteomics combined to advanced bioinformatics tools is acquiring a pivotal role in the comprehensive understanding of living organism's biology, in particular for non-model organisms, which includes most marine and aquatic invertebrates. Depicting of protein composition in a whole organ/organism followed by their assembling in functional protein association networks promotes the understanding of key biological processes. Here, we provide a detailed description of the extraction procedure of cell-free coelomic fluid soluble proteins and the characterization of the proteome of the starfish Marthasterias glacialis. Due to coelomic fluid richness in glycoproteins, which complicates protein identification, extracts of soluble proteins are deglycosylated prior to tandem mass spectrometry. This experimental approach is useful at improving knowledge on the coelomic fluid physiological role and deciphering its involvement in regeneration of starfish body parts when comparing different regeneration conditions.
蛋白质组学与先进的生物信息学工具相结合，在全面理解生物体生物学方面正发挥着关键作用，特别是对于非模式生物，其中包括大多数海洋和水生无脊椎动物。描绘整个器官/生物体中的蛋白质组成，然后将它们组装到功能性蛋白质关联网络中，有助于理解关键的生物过程。在这里，我们详细描述了无细胞体腔液可溶性蛋白质的提取过程以及海星Marthasterias glacialis蛋白质组的特征。由于体腔液中糖蛋白丰富，这使得蛋白质鉴定变得复杂，因此在串联质谱分析之前，先对可溶性蛋白质提取物进行去糖基化处理。当比较不同的再生条件时，这种实验方法有助于提高对体腔液生理作用的认识，并解读其在海星身体部位再生中的作用。
