Spatial and temporal regulation of protein kinase D (PKD)

EMBO J. 2000 Jun 15;19(12):2935-45. doi: 10.1093/emboj/19.12.2935.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • B-Lymphocytes / enzymology*
  • Binding Sites
  • Biological Transport
  • Cell Compartmentation
  • Cell Membrane / enzymology*
  • Cell Membrane / ultrastructure
  • Diglycerides / metabolism
  • Gene Expression Regulation, Enzymologic
  • Glycosphingolipids
  • Green Fluorescent Proteins
  • Luminescent Proteins / genetics
  • Luminescent Proteins / isolation & purification
  • Luminescent Proteins / metabolism
  • Mast Cells / enzymology*
  • Protein Kinase C / genetics
  • Protein Kinase C / isolation & purification
  • Protein Kinase C / metabolism*
  • Receptors, Antigen, B-Cell / metabolism*
  • Recombinant Fusion Proteins / isolation & purification
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction

Substances

  • Diglycerides
  • Glycosphingolipids
  • Luminescent Proteins
  • Receptors, Antigen, B-Cell
  • Recombinant Fusion Proteins
  • Green Fluorescent Proteins
  • protein kinase D
  • Protein Kinase C