School of Life Sciences, the Key Laboratory of Plant Cell Engineering and Germplasm Innovation, Ministry of Education, Shandong University, Jinan, 250100, PR China.
BMC Genomics. 2009 Dec 12;10:600. doi: 10.1186/1471-2164-10-600.
Metamorphosis is an important process in the life cycle of holometabolous insects and is regulated by insect hormones. During metamorphosis, the epidermis goes through a significant transformation at the biochemical and molecular levels.
To identify proteins and phosphoproteins involved in this process, we separated and compared epidermal protein profiles between feeding larvae and metamorphically committed larvae using two-dimensional gel electrophoresis and Pro-Q Diamond Phosphoprotein Staining. Sixty-one spots showing differential expression and/or phosphorylation were analyzed by mass spectrometry and eighteen proteins were proved related to larval-pupal transformation. Eight of them were further examined at the mRNA level by Reverse Transcription Polymerase Chain Reaction (RT-PCR) and two of them were examined at the protein level by Western blot. Calponin was highly expressed in the metamorphic epidermis and phosphorylated by protein kinase C.
Our results suggest that the expression and phosphorylation of these proteins may play important roles in coordinating the biochemical processes involved in larval-pupal metamorphosis.
变态是完全变态昆虫生命周期中的一个重要过程，由昆虫激素调控。在变态过程中，表皮在生化和分子水平上经历显著的转化。
为了鉴定参与这一过程的蛋白质和磷酸化蛋白质，我们使用二维凝胶电泳和 Pro-Q Diamond Phosphoprotein Staining 分离和比较了取食幼虫和变态期幼虫的表皮蛋白图谱。通过质谱分析鉴定了 61 个显示差异表达和/或磷酸化的斑点，并用反转录聚合酶链反应 (RT-PCR) 在 mRNA 水平进一步检测了其中的 18 种蛋白质，用 Western blot 检测了其中的 2 种蛋白质。钙调蛋白在变态表皮中高表达，并被蛋白激酶 C 磷酸化。
我们的结果表明，这些蛋白质的表达和磷酸化可能在协调幼虫-蛹变态过程中涉及的生化过程中发挥重要作用。
