Solution and ion-complexed conformations of beauvericin determined by proton magnetic resonance spectroscopy

Abstract

The conformational properties of the cyclohexadepsipeptide antibiotic Beauvericin have been investigated by 1H-NMR spectroscopy in polar (C 2H 3O 2H) and non-polar (CCI 4, C 6 2H 6, C 2HCI 3) solvents and in two solvent mixtures; one a mixture of a polar and non-polar solvent (C 2H 3O 2H/CCI 4) and the other an aromatic solvent in a non-polar environment (C 6 2H 6/CCI 4). The ion-complexation properties of Beauvericin with alkali metal halides (Li +, Na +, K +, Cs +) have also been studied. It is demonstrated that changes in chemical shifts of Beauvericin with concentration, with polarity of solvent or with added alkali metal ion reflect changes not only in the solvent properties but also changes in backbone conformation and changes due to ion-complexation, where appropriate, and therefore cannot be used, by themselves, to determine the conformation of the molecule, its self-aggregation properties, or the stoichiometry of the metal ion-co mplex. The backbone conformations of Beauvericin in different environments are determined by methods that are independent of chemical shift analysis; i.e., by measurements of 5 J(HH) magnitudes observed between the α-CH protons of the l-phenylalanine and d-hydroxyisovaleric acid ( dHyIv) residues and by nuclear Overhauser effect measurements observed between α-CH(HyIv) and ( N)-CH 3(Phe) proton signals. In the knowledge of these results the chemical shifts of Beauvericin in different environments can then be rationalised. It is found that the conformation of Beauvericin in a polar solvent is different from that found in a non-polar solvent and from that found for the ion-complexed form in a polar solvent; all the evidence available suggests that the conformation in the ion-complexed form is similar to that found in non-polar solvents. By taking into account the conformational properties of the l-phenylalanine and dHyIv side-chains, it is possible to assign unambiguously the magnetically non-equivalent β-CH 2(Phe) and γMe(HyIv) proton signals and so elucidate the complete conformational behaviour of the uncomplexed forms of Beauvericin in a polar and a non-polar environment, and of the ion-complexed form of Beauvericin in a polar solvent.