The interactions of the brain-specific calmodulin-binding protein kinase C substrate, neuromodulin (GAP 43), with membrane phospholipids.
The interaction of the brain-specific calmodulin-binding protein kinase C (PKC) substrate, neuromodulin (GAP 43), with membrane phospholipids was studied. Specific binding of neuromodulin to negatively charged phospholipids through electrostatic interactions was demonstrated. Comparison of the binding of neuromodulin to acidic phospholipids with that of neurogranin, a newly characterized calmodulin-binding PKC substrate (Baudier J., Deloulme, J. C., Van Dorsselaer, A., Black, D., and Mathes H. (1991) J. Biol. Chem. 266, 229-237) suggested that the conserved basic amino acid sequence which characterizes the two proteins and which corresponds to the PKC phosphorylation and calmodulin binding domain also serves as phospholipid binding site. In the absence of calmodulin, binding of neuromodulin to phosphatidylserine at low concentration parallels its phosphorylation by PKC, suggesting that formation of a ternary complex between neuromodulin, phosphatidylserine, and PKC is required for optimum neuromodulin phosphorylation. In the presence of calmodulin, the binding of neuromodulin to phosphatidylserine is inhibited, resulting in total inhibition of neuromodulin phosphorylation. Our results suggest that, in vivo, phosphorylation of neuromodulin may not only depend on protein kinase C (PKC)1 activation but also on the accessibility of the neuromodulin phosphorylation domain to activated membrane-bound PKC that could regulated by CaM.
Houbre
D
D
Laboratoire de Biophysique, Centre de Recherches Pharmaceutiques, URA 491 du Centre National de la Recherche Scientifique, Université Louis Pasteur, Illkirch, France.
Duportail
G
G
Deloulme
J C
JC
Baudier
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002148:Calmodulin-Binding Proteins / Q000378:metabolism*; D002417:Cattle; D003600:Cytosol / Q000378:metabolism; D019922:GAP-43 Protein; D007700:Kinetics; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism*; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D051579:Neurogranin; D010743:Phospholipids / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D011485:Protein Binding; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D013816:Thermodynamics
D002148:Calmodulin-Binding Proteins; D019922:GAP-43 Protein; D008562:Membrane Glycoproteins; D008563:Membrane Lipids; D009419:Nerve Tissue Proteins; C496651:Nrgn protein, rat; D010743:Phospholipids; D010750:Phosphoproteins; D051579:Neurogranin; D011493:Protein Kinase C