Molly Mason and Peter Kirkpatrick

Abstract:

GRK2 regulates G protein signaling pathways and p38 is a member of the mitogen activated protein kinase (MAPK) family involved in transcriptional response to cellular stresses. These two pathways are intertwined as GRK2 phosphorylates and inhibits p38. However, details of the GRK2-p38 interaction remain unknown, especially what domains of GRK2 are required for binding p38. We developed an in vitro assay that measures full-length GRK2 binding to GST-p38 immobilized on glutathione agarose beads. We recognized that if we could immobilize GRK2 we could use the glutathione-S-transferase activity of GST-p38 to quantify binding. Our current goals are 1) to see if we can reproduce GST-p38 binding to GRK2 immobilized on nickel beads and 2) to determine what domains of GRK2 are involved in p38 binding.