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Investigation into the mechanism regulating MRP localization

Iman van den Bout1, Jacco van Rheenen, Annelies A van Angelen

  • 1Division of Cell Biology, Netherlands Cancer Institute, 121 Plesmanlaan, 1066 CX Amsterdam, The Netherlands.

Experimental Cell Research
|September 28, 2007
PubMed

Insights

MacMARCKS (MRP) protein localization to the plasma membrane requires its effector domain (ED) but not Ptdins(4,5)P2 binding. This highlights a novel Ptdins(4,5)P2-independent mechanism for membrane targeting in cellular processes.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Major protein kinase C (PKC) substrates MARCKS and MacMARCKS (MRP) are involved in cell spreading, integrin activation, and exocytosis.
  • The myristoyl-electrostatic switch model explains membrane binding through myristoyl moiety and effector domain (ED) interaction, with electrostatic loss causing cytoplasmic translocation.

Purpose of the Study:

  • To investigate the mechanisms regulating MacMARCKS (MRP) localization.
  • To determine the role of the effector domain (ED) and Ptdins(4,5)P2 in MRP membrane binding and localization.

Main Methods:

  • Phosphorylation of MRP and subsequent analysis of its subcellular localization.
  • In vitro binding assays to assess the interaction of MRP's ED with phospholipids.

Main Results:

  • Phosphorylated MRP is relocated to intracellular membranes of late endosomes and lysosomes.
  • MRP binds to all membranes via its myristoyl moiety; however, ED is crucial for plasma membrane localization.
  • While MRP's ED binds Ptdins(4,5)P2 in vitro, this interaction is not essential for plasma membrane targeting or retention.

Conclusions:

  • The co-operation between the myristoyl moiety and ED is essential for targeting MRP to the plasma membrane.
  • MRP's plasma membrane targeting occurs in a Ptdins(4,5)P2-independent manner, differing from the classical myristoyl-electrostatic switch model for MARCKS.

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