Tay Tuan Leng, Lin Qingsong, Seow Teck Keong, Tan Keng Hwa, Hew Choy Leong, Gong Zhiyuan
Department of Biological Sciences, National University of Singapore, Singapore 119260.
Proteomics. 2006 May;6(10):3176-88. doi: 10.1002/pmic.200600030.
In the present study, profiles of protein expression were examined during early development of zebrafish, an increasingly popular experimental model in vertebrate development and human diseases. By 2-DE, an initial increase in protein spots from 6 h post-fertilization (hpf) to 8-10 hpf was observed. There was no dramatic change in protein profiles up to 18 hpf, but significant changes occurred in subsequent stages. Interestingly, 49% of the proteins detected at 6 hpf remained detectable by 1 week of age. To map the protein expression patterns in 2-D gels, MALDI-TOF/TOF MS was employed to identify selected protein spots from early embryos. 108 protein spots were found to match known proteins and they were derived from 55 distinct genes. Interestingly, 11 (20%) of them produced multiple protein isoforms or distinct cleavage products. Although deyolked embryos were used in the analysis, a large number of vitellogenin derivatives remained prominently present in the embryos. Other than these, most of the identified proteins are cytosolic, cytoskeletal and nuclear proteins, which are involved in diversified functions such as metabolism, cytoskeleton, translation, protein degradation, etc. Some of the proteins with interesting temporal expression profiles during development are further discussed.
在本研究中,我们检测了斑马鱼早期发育过程中的蛋白质表达谱。斑马鱼是脊椎动物发育和人类疾病研究中越来越受欢迎的实验模型。通过二维电泳(2-DE),我们观察到从受精后6小时(hpf)到8-10 hpf蛋白质斑点数量初步增加。直到18 hpf蛋白质谱都没有显著变化,但在随后的阶段发生了显著变化。有趣的是,在6 hpf检测到的蛋白质中有49%在1周龄时仍可检测到。为了在二维凝胶中绘制蛋白质表达模式,我们采用基质辅助激光解吸/电离飞行时间串联质谱(MALDI-TOF/TOF MS)来鉴定早期胚胎中选定的蛋白质斑点。发现108个蛋白质斑点与已知蛋白质匹配,它们来自55个不同的基因。有趣的是,其中11个(20%)产生多种蛋白质异构体或不同的裂解产物。尽管分析中使用的是去卵黄胚胎,但大量卵黄蛋白原衍生物在胚胎中仍然显著存在。除此之外,大多数鉴定出的蛋白质是胞质、细胞骨架和核蛋白,它们参与多种功能,如代谢、细胞骨架、翻译、蛋白质降解等。我们进一步讨论了一些在发育过程中具有有趣时间表达谱的蛋白质。