SEARCH

SEARCH BY CITATION

Abstract

The all-light-atom X-ray crystal-structure analysis of cyclosporin A (1), a cyclic undecapeptide containing seven N-methylated amino acids, reveals a conformation very similar to that of the previously analysed iodo derivative which is characterised by a twisted β-pleated sheet involving the residues Me-Val-11, MeBmt-1, Abu-2, Sar-3, MeLeu-4, Val-5, MeLeu-6, and Ala-7. The β-bend at Sar-MeLeu-4 is of type II′, and the loop of the residual amino acids involves a cis-peptide bond between MeLeu-9 and MeLeu-10. The NH proton of D-Ala-8 closes a yi-bend with a H-bond to the MeLeu-6 CO group. The crystal was grown from acetone. A closely similar backbone conformation in apolar solvents such as CDCl3 or C6D6 has been derived from the interpretation of the NMR spectral parameters (homo- and heteronuclear NOE effects, coupling constants, chemical-shift values of the C-, H-, and N-atoms). A minor variation in the backbone conformation between crystal and solution is observed in the region of D-Ala-8, where in solution a 3-center H-bond is established between the NH of D-Ala-8 und the carbonyl O-atoms of both MeLeu-6 (yi-turn) and D-Ala-8 (C5-bend). A recently proposed technique to identify intramolecular H-bond via heteronuclear NOE from NH proton to carbonyl C-atoms is critically analysed. The main difference between crystal and solution conformations lies in the orientation of the side chains of the unusual amino acid MeBmt (χ1 = +60° in solution, -168° in the crystal) and of MeLeu-10 (X1 = −60° in solution, +60° in the crystal). The differences in crystal and solution are caused by the break of the intermolecular H-bond of the OH group of MeBmt on dissolution of the crystal. The bifurcated H-bond of D-Ala-8 twists the backbone in this region. Molecular modeling demonstrates that this is the origin of the change in the side chain conformation of MeLeu-10. The intramolecular flexibility in the crystal indicated by the thermal parameters obtained from the X-ray refinement, and in solution by an analysis of spin-lattice relaxation times in the NMR experiments, indicate a fairly rigid backbone and fixed conformations for all the side chains except for that of Abu-2 and the distal atoms of MeBmt.