The inner leaflet of a typical mammalian plasma membrane contains 20-30% univalent PS (phosphatidylserine) and 1% multivalent PtdIns(4,5)P(2). Numerous proteins have clusters of basic (or basic/hydrophobic) residues that bind to these acidic lipids. The intracellular effector CaM (calmodulin) can reverse this binding on a wide variety of proteins, including MARCKS (myristoylated alanine-rich C kinase substrate), GAP43 (growth-associated protein 43, also known as neuromodulin), gravin, GRK5 (G-protein-coupled receptor kinase 5), the NMDA (N-methyl-D-aspartate) receptor and the ErbB family. We used the first principles of physics, incorporating atomic models and the Poisson-Boltzmann equation, to describe how the basic effector domain of MARCKS binds electrostatically to acidic lipids on the plasma membrane. The theoretical calculations show the basic cluster produces a local positive electrostatic potential that should laterally sequester PtdIns(4,5)P(2), even when univalent acidic lipids are present at a physiologically relevant 100-fold excess; four independent experimental measurements confirm this prediction. Ca(2+)/CaM binds with high affinity (K(d) approximately 10nM) to this domain and releases the PtdIns(4,5)P(2). MARCKS, a major PKC (protein kinase C) substrate, is present at concentrations comparable with those of PtdIns(4,5)P(2) (approx. 10 microM) in many cell types. Thus MARCKS can act as a reversible PtdIns(4,5)P(2) buffer, binding PtdIns(4,5)P(2) in a quiescent cell, and releasing it locally when the intracellular Ca(2+) concentration increases. This reversible sequestration is important because PtdIns(4,5)P(2) plays many roles in cell biology. Less is known about the role of CaM-mediated reversible membrane binding of basic/hydrophobic clusters for the other proteins.
[1]
D. Murray,et al.
Electrostatic sequestration of PIP2 on phospholipid membranes by basic/aromatic regions of proteins.
,
2004,
Biophysical journal.
[2]
Diana Murray,et al.
A computational model for the electrostatic sequestration of PI(4,5)P2 by membrane-adsorbed basic peptides.
,
2004,
Biophysical journal.
[3]
Diana Murray,et al.
Genome-wide analysis of membrane targeting by S. cerevisiae pleckstrin homology domains.
,
2004,
Molecular cell.
[4]
B Honig,et al.
Binding of small basic peptides to membranes containing acidic lipids: theoretical models and experimental results.
,
1996,
Biophysical journal.
[5]
H. Stenmark.
Phosphoinositides in Subcellular Targeting and Enzyme Activation
,
2004,
Current Topics in Microbiology and Immunology.
[6]
H. Helmholtz,et al.
Two monographs on electrokinetics
,
1951
.