Calmodulin interacts with amphiphilic peptides composed of all D-amino acids.
Calmodulin binds to amphiphilic, helical peptides of a variety of amino-acid sequences. These peptides are usually positively charged, although there is spectroscopic evidence that at least one neutral peptide binds. The complex between calmodulin and one of its natural target peptides, the binding site for calmodulin on smooth muscle myosin light-chain kinase (RS20), has been investigated by crystallography and NMR which have characterized the interactions between the ligand and the protein. From these data, it appears that the calmodulin-binding surface is sterically malleable and van der Waals forces probably dominate the binding. To explore further this apparently permissive binding, we investigated the chiral selectivity of calmodulin using synthesized analogues of melittin and RS20 that consisted of only D-amino acids. Fluorescence and NMR measurements show that D-melittin and D-RS20 both bind avidly to calmodulin, probably in the same general binding site as that for peptides having all L-amino acids. The calmodulin-peptide binding surface is therefore remarkably tolerant sterically. Our results suggest a potentially useful approach to the design of non-hydrolysable or slowly hydrolysable intracellular inhibitors of calmodulin.
Fisher
P J
PJ
Department of Pharmacology, Mayo Clinic, Rochester, Minnesota 55905.
Prendergast
F G
FG
Ehrhardt
M R
MR
Urbauer
J L
JL
Wand
A J
AJ
Sedarous
S S
SS
McCormick
D J
DJ
Buckley
P J
PJ
D001665:Binding Sites; D002147:Calmodulin / Q000378:metabolism*; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D002942:Circular Dichroism; D005454:Fluorescence Polarization; D008555:Melitten / Q000378:metabolism; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D011485:Protein Binding; D013056:Spectrophotometry, Ultraviolet; D013237:Stereoisomerism
D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D010455:Peptides; C061167:RS 20; D008555:Melitten; D009219:Myosin-Light-Chain Kinase