Submitted to: In Vitro Biology Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 4/15/2013
Publication Date: N/A
Technical Abstract: Prostaglandins (PGs) play crucial roles in vertebrate biology, particularly in immune functions. Because PGs also mediate specific cell functions in insect immunity, we are investigating how these signaling molecules affect insect cells. We reported that PGs, notably PGA1, PGA2, and PGE1, up and/or down regulate expression of proteins involved in cellular protection, signal transduction, protein structure/degradation, and cell metabolism/energetics. Here we report on how they influence protein phosphorylation, an important action in intracellular signal transduction. We incubated the corn earworm cell line, BCIRL-HzAM1, with 15 uM PGA2, PGE1 or PGF2a'for 20 min or 40 min and then subjected the resulting homogenates to 2D electrophoresis. We stained the gels with a phosphoprotein-specific stain, as well as a total protein stain. Following the 20 min incubations, we noted changes in the phosphorylation of 7 proteins incubated with PGA2, 10 proteins with PGE1, and 8 proteins with PGF2a. We analyzed the proteins using MS/MS (MALDI TOF/TOF) and, with bioinformatics, determined their functions. For PGA2, phosphorylation increased in 2 proteins involved in protein-protein interactions, 1 in transcription, 1 in RNA processing; phosphorylation decreased in 2 proteins involved in protein degradation/folding and 1 in metabolism. For PGE1, phosphorylation increased in 5 proteins involved in protein folding/degradation and 2 in cell structure or metabolism; phosphorylation decreased for 3 proteins involved in cell proliferation, signal transduction and RNA processing. For PGF2a, phosphorylation increased in 1 protein involved in protein folding and another in cell cycle control; phosphorylation decreased in 5 proteins involved in protein folding/degradation, protein synthesis or protein-protein interactions, and increased in one protein associated with metabolism. We are determining changes in protein phosphorylation for cells exposed to PGs for 40 min and will report our results.