title: Structural changes in melittin and calmodulin upon complex formation and their modulation by calcium.
abstract: In the presence of Ca2+, calmodulin forms a 1:1 high-affinity complex (Kd = 3 nM) with melittin, a peptide from bee venom; in the presence of ethylenediaminetetraacetic acid, a second type of complex, of much lower affinity, is formed [Comte, M., Maulet, Y., & Cox, J. A. (1983) Biochem. J. 209, 269-272]. In this paper, these interactions were studied by tryptophan fluorescence and circular dichroism spectroscopy in near- and far-UV. Interaction between the two peptides in the presence as well as in the absence of Ca2+ leads to the shielding of the tryptophan residue of melittin from its aqueous environment and to an increase in the alpha-helical content of bound melittin; for instance the Ca2+-dependent high-affinity complex formation enhances the alpha-helical content of melittin from 5 to 72%. Provided Ca2+ is present, the interaction between the two peptides leads to significant changes in the environment of at least one tyrosine residue of calmodulin as measured by near-UV circular dichroism. In the absence of Ca2+, calmodulin binds two melittin molecules with a Kd of ca. 10 microM; at higher concentrations of free melittin, additional binding occurs (up to 5 mol of melittin/mol of calmodulin), with concomitant denaturation of calmodulin. In the presence of 4.0 M urea, the low-affinity complexes formed in the absence of Ca2+ dissociate, due to the denaturation of metal-free calmodulin, whereas the spectroscopic signals of the high-affinity Ca2+-dependent complex are not affected.(ABSTRACT TRUNCATED AT 250 WORDS)
mesh_terms: D000818:Animals; D001514:Bee Venoms / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D002417:Cattle; D002942:Circular Dichroism; D007700:Kinetics; D008555:Melitten / Q000378:metabolism*; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence
chemical_list: D001514:Bee Venoms; D002147:Calmodulin; D008555:Melitten; D002118:Calcium