Modelling and refinement of the conformation of mycosubtilin in solution from two-dimensional NMR data.

The conformation in solution of mycosubtilin, an antifungal lipopeptide [cyclo(L-Asn-D-Tyr-D-Asn-L-Gln-L-Pro-D-Asn-L-Ser-beta-amino acid)]has been probed by two-dimensional nuclear magnetic resonance and restrained energy minimization. Several structures have been proposed belonging to the same family with minor local variations related to different orientations of amide planes. The molecular topology was found to be completely different from that of iturin A, an analogue which exhibits quite different biological properties. The cyclic peptide of mycosubtilin is shown to be rather rigid in the region of L-proline and stabilized by C7 structures; in contrast, the neighbourhood of D-tyrosine-2 was found to be more flexible. The validity of our models is discussed first in terms of distance violation and second on the basis of reconstructed NOE spectroscopy maps. The different limitations towards higher-resolution structures are discussed.