Subramanian Perumal, Jayapalan Jaime J, Abdul-Rahman Puteri S, Arumugam Manjula, Hashim Onn H
Department of Biochemistry and Biotechnology, Annamalai University , Chidambaram, Tamil Nadu , India.
University of Malaya Centre for Proteomics Research (UMCPR), Faculty of Medicine, University of Malaya , Kuala Lumpur , Malaysia.
PeerJ. 2016 May 24;4:e2080. doi: 10.7717/peerj.2080. eCollection 2016.
Background. Diurnal rhythms of protein synthesis controlled by the biological clock underlie the rhythmic physiology in the fruit fly, Drosophila melanogaster. In this study, we conducted a proteome-wide investigation of rhythmic protein accumulation in D. melanogaster. Materials and Methods. Total protein collected from fly samples harvested at 4 h intervals over the 24 h period were subjected to two-dimensional gel electrophoresis, trypsin digestion and MS/MS analysis. Protein spots/clusters were identified with MASCOT search engine and Swiss-Prot database. Expression of proteins was documented as percentage of volume contribution using the Image Master 2D Platinum software. Results. A total of 124 protein spots/clusters were identified using MS/MS analysis. Significant variation in the expression of 88 proteins over the 24-h period was observed. A relatively higher number of proteins was upregulated during the night compared to the daytime. The complexity of temporal regulation of the D. melanogaster proteome was further reflected from functional annotations of the differently expressed proteins, with those that were upregulated at night being restricted to the heat shock proteins and proteins involved in metabolism, muscle activity, protein synthesis/folding/degradation and apoptosis, whilst those that were overexpressed in the daytime were apparently involved in metabolism, muscle activity, ion-channel/cellular transport, protein synthesis/folding/degradation, redox homeostasis, development and transcription. Conclusion. Our data suggests that a wide range of proteins synthesized by the fruit fly, D. melanogaster, is under the regulation of the biological clock.
背景。生物钟控制的蛋白质合成昼夜节律是果蝇（黑腹果蝇）节律性生理的基础。在本研究中，我们对黑腹果蝇中节律性蛋白质积累进行了全蛋白质组研究。材料与方法。在24小时内每隔4小时采集的果蝇样本中提取的总蛋白质，进行二维凝胶电泳、胰蛋白酶消化和串联质谱分析。蛋白质斑点/簇用MASCOT搜索引擎和Swiss-Prot数据库进行鉴定。使用Image Master 2D Platinum软件将蛋白质表达记录为体积贡献百分比。结果。通过串联质谱分析共鉴定出124个蛋白质斑点/簇。观察到88种蛋白质在24小时内的表达有显著变化。与白天相比，夜间上调的蛋白质数量相对较多。黑腹果蝇蛋白质组时间调控的复杂性从差异表达蛋白质的功能注释中进一步体现出来，夜间上调的蛋白质局限于热休克蛋白以及参与代谢、肌肉活动、蛋白质合成/折叠/降解和细胞凋亡的蛋白质，而白天过表达的蛋白质显然参与代谢、肌肉活动、离子通道/细胞运输、蛋白质合成/折叠/降解、氧化还原稳态、发育和转录。结论。我们的数据表明，果蝇（黑腹果蝇）合成的多种蛋白质受生物钟调控。
