前列腺素(PG)是花生四烯酸和其他两种C20多不饱和脂肪酸的氧化代谢产物。在无脊椎动物的其他行为中,PGs在卵巢发育中起作用,肾功能,豁免权,血细胞迁移,和基因/蛋白质表达。可逆磷酸化是调节真核细胞中蛋白质功能的主要机制,对于某些哺乳动物蛋白质,它受到PG的影响。我们提出了一个假设,即PGs影响昆虫细胞内的蛋白质磷酸化,我们用已建立的昆虫细胞系进行了测试,BCIRL-HzAM1。在三个PG之一(15μM)的存在下孵育20、30或40分钟后,PGA2,PGE1,或PGF2α,对不同的细胞组进行双向电泳和串联质谱分析。我们记录了31种蛋白质的显著磷酸化变化,15中减少,15中增加,一种磷酸化增加或减少的蛋白质,取决于PG治疗。增加PG暴露时间导致更少的蛋白质变化,20分钟的孵育导致16种蛋白质的变化,30分钟到13的变化,40分钟到2蛋白质的变化。通过生物信息学分析鉴定了蛋白质,包括成绩单描述,计算的分子量和等电点,MOlecularWeightSEarch评分,总离子分数,肽的数量,蛋白质覆盖率百分比,E值,和最高的肽得分。本文提供的数据坚定地支持了我们的假设,即PG会影响昆虫细胞内的蛋白质磷酸化,并为昆虫生物学增加了一种新的PG信号功能。
Prostaglandins (PGs) are oxygenated metabolites of arachidonic acid and two other C20 polyunsaturated fatty acids. Among other actions in invertebrates, PGs act in ovarian development, renal functions, immunity, hemocyte migration, and gene/protein expression. Reversible phosphorylation is a major mechanism of regulating protein functions in eukaryotic cells and for some mammalian proteins it is influenced by PGs. We posed the hypothesis that PGs influence protein phosphorylation within insect cells, which we tested with the established insect cell line, BCIRL-HzAM1. After 20, 30, or 40 min incubations in the presence of one of three PGs (at 15 μM), PGA2 , PGE1 , or PGF2α , separate sets of cells were processed for analysis by two-dimensional electrophoresis followed by tandem mass spectrometry. We recorded significant phosphorylation changes in 31 proteins, decreases in 15, and increases in 15, and one protein with increased or decreased phosphorylation, depending on PG treatment. Increasing PG exposure times led to changes in fewer proteins, 20 min incubations led to changes in 16 proteins, 30 min to changes in 13, and 40 min to changes in 2 proteins. The proteins were identified by bioinformatic analyses, including transcript description, calculated molecular weights and isoelectric points, MOlecular Weight SEarch score, total ion score, numbers of peptides, percent protein coverage, E-value, and highest peptide score. The data presented in this paper firmly support our hypothesis that PGs influence protein phosphorylation within insect cells and adds a novel PG-signaled function to insect biology.