Prostaglandins influence protein phosphorylation in established insect cell line

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 PGF_2α , 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.