Solid-state NMR conformational studies of a melittin-inhibitor complex.
Melittin is a cytolytic peptide whose biological activity is lost upon binding to a six-residue peptide, Ac-IVIFDC-NH(2), with which it forms a highly insoluble complex. As a result, the structural analysis of the interaction between the two peptides is difficult. Solid-state NMR spectroscopy was used to study the interaction between melittin and the peptide inhibitor. Location of the binding site in the melittin-inhibitor complex was determined using lanthanide ions, which quench NMR resonances from molecular sites that are in close proximity to the unique ion binding site. Our results indicated that the inhibitor binding site in melittin is near Leu13, Leu16 and Ile17, but not near Leu6 or Val8. On the basis of these data we propose that the inhibitor binds to melittin in the vicinity of Ala15 to Trp19 and prevents insertion of melittin into cell membranes by disrupting the helical structure. Supporting evidence for this model was produced by determining the distance, using rotational resonance NMR, between the [1-(13)C] of Leu13 in melittin and the [3-(13)C] of Phe4 in the inhibitor.
Lam
Y-H
YH
School of Chemistry, University of Melbourne, Parkville, Victoria 3010, Australia.
Morton
C J
CJ
Separovic
F
F
D000123:Acetyltransferases; D051046:Amino-Acid N-Acetyltransferase; D001665:Binding Sites; D028581:Lanthanoid Series Elements / Q000737:chemistry; D046911:Macromolecular Substances; D008555:Melitten / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009842:Oligopeptides / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D025941:Protein Interaction Mapping / Q000379:methods*
C474040:Ac-IVIFDC-NH(2); D028581:Lanthanoid Series Elements; D046911:Macromolecular Substances; D009842:Oligopeptides; D010455:Peptides; D008555:Melitten; D000123:Acetyltransferases; D051046:Amino-Acid N-Acetyltransferase