Spatial and temporal regulation of protein kinase D (PKD).
Protein kinase D (PKD; also known as PKCmicro) is a serine/threonine kinase activated by diacylglycerol signalling pathways in a variety of cells. PKD has been described previously as Golgi-localized, but herein we show that it is present within the cytosol of quiescent B cells and mast cells and moves rapidly to the plasma membrane after antigen receptor triggering. The membrane redistribution of PKD requires the diacylglycerol-binding domain of the enzyme, but is independent of its catalytic activity and does not require the integrity of the pleckstrin homology domain. Antigen receptor signalling initiates in glycosphingolipid-enriched microdomains, but membrane-associated PKD does not co-localize with these specialized structures. Membrane targeting of PKD is transient, the enzyme returns to the cytosol within 10 min of antigen receptor engagement. Strikingly, the membrane-recycled PKD remains active in the cytosol for several hours. The present work thus characterizes a sustained antigen receptor-induced signal transduction pathway and establishes PKD as a serine kinase that temporally and spatially disseminates antigen receptor signals away from the plasma membrane into the cytosol.
Matthews
S A
SA
Lymphocyte Activation Laboratory and Molecular Neuropathobiology Laboratory, Imperial Cancer Research Fund, 44 Lincoln's Inn Fields, London WC2A 3PX, UK.
Iglesias
T
T
Rozengurt
E
E
Cantrell
D
D
D001402:B-Lymphocytes / Q000201:enzymology*; D001665:Binding Sites; D001692:Biological Transport; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000201:enzymology* / Q000648:ultrastructure; D004075:Diglycerides / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D006028:Glycosphingolipids; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008407:Mast Cells / Q000201:enzymology*; D011493:Protein Kinase C / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D015398:Signal Transduction
D004075:Diglycerides; D006028:Glycosphingolipids; D008164:Luminescent Proteins; D011947:Receptors, Antigen, B-Cell; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; C089212:protein kinase D; D011493:Protein Kinase C