Division of Life Science, Hong Kong University of Science and Technology, Hong Kong SAR, China.
PLoS One. 2012;7(6):e38814. doi: 10.1371/journal.pone.0038814. Epub 2012 Jun 15.
The polychaete Neanthes arenaceodentata- is cosmopolitan in distribution-, has been used as a laboratory test animal. Life history of this species has several unique features; the female dies after spawning and the male incubates the fertilized eggs through the 21-segmented stage. The larvae leave the tube and commence feeding. Changes in protein abundance and phosphorylation were examined during early development of N. arenaceodentata. A gel-based approach and gel-free enrichment of phosphopeptides coupled with mass spectrometry were used to identify proteins and phosphopeptides in fertilized ova and larval stages. Patterns of proteins and phosphoproteins changed from fertilized ova to larval stages. Twelve proteins occurred in phosphorylated form and nine as stage specific proteins. Cytoskeletal proteins have exhibited differential phosphorylation from ova to larval stages; whereas, other proteins exhibited stage-specific phosphorylation patterns. Ten phosphopeptides were identified that showed phosphorylation sites on serine or threonine residues. Sixty percent of the identified proteins were related to structural reorganization and others with protein synthesis, stress response and attachment. The abundance and distribution of two cytoskeleton proteins were examined further by 2-DE Western blot analysis. This is the first report on changes in protein expression and phosphorylation sites at Thr/Ser in early development of N. arenaceodentata. The 2-DE proteome maps and identified phosphoproteins contributes toward understanding the state of fertilized ova and early larval stages and serves as a basis for further studies on proteomics changes under different developmental conditions in this and other polychaete species.
沙蚕 Neanthes arenaceodentata-分布广泛，已被用作实验室测试动物。该物种的生活史具有几个独特的特征；雌性在产卵后死亡，雄性通过 21 个节段孵化受精卵。幼虫离开管并开始进食。本研究采用凝胶电泳和无凝胶磷酸肽富集联合质谱技术，研究了沙蚕早期发育过程中蛋白质丰度和磷酸化的变化。从受精卵到幼虫阶段，蛋白质和磷酸化蛋白质的模式发生了变化。有 12 种蛋白质以磷酸化形式存在，9 种蛋白质为阶段特异性蛋白质。从受精卵到幼虫阶段，细胞骨架蛋白表现出不同的磷酸化；而其他蛋白质则表现出特定阶段的磷酸化模式。鉴定出 10 个磷酸肽，其丝氨酸或苏氨酸残基上有磷酸化位点。所鉴定的蛋白质中有 60%与结构重排有关，其他蛋白质与蛋白质合成、应激反应和附着有关。通过 2-DE Western blot 分析进一步研究了两种细胞骨架蛋白的丰度和分布。这是首次报道沙蚕早期发育过程中 Thr/Ser 磷酸化位点的蛋白质表达和磷酸化变化。2-DE 蛋白质组图谱和鉴定的磷酸化蛋白有助于了解受精卵和早期幼虫阶段的状态，并为在该物种和其他多毛类动物中研究不同发育条件下蛋白质组变化提供了基础。
