Conformational change of ascidiacyclamide caused by asymmetric modification for an isoleucine residue: structural analyses of [Gly], [Leu], and [Phe]ascidiacyclamides by x-ray diffraction and NMR spectroscopy.
Ascidiacyclamide, a cytotoxic cyclic peptide from tunicate, is composed of unusual amino acids and has a repeated sequence, c[-thiazole-D-Val-oxazoline-L-Ile-]2 ([Ile]ASC). The symmetric chemical structure has been assumed to be correlated with the cytotoxicity, and it is reasonable to consider that the disturbance of its structure from the C2 symmetry results in the changes of conformation and activity. In order to quantitatively estimate the molecular conformation-activity relationship, an isoleucine residue was substituted by Gly, Leu, or Phe to disturb the C2 symmetry. The conformations of three derivatives were examined by nmr spectroscopy and the crystal structure of [Leu]ASC was also analyzed by x-ray diffraction method. The 1H-nmr experiments and the constrained molecular dynamics simulations showed the twisted "figure 8" conformers for [Gly] and [Phe]ASCs and the "square" conformer for [Leu]ASC in the DMSO solution. The x-ray crystal analysis of [Leu]ASC also revealed the square form similar to the solution structure. On the other hand, their cytotoxic activities were measured using L1210 leukemia cells and were related with the bulkiness and/or hydrophobicity of the side chain of the substituted amino acid; [Phe] > or = [Ile] > [Leu] >> [Gly]ASCs. As an attempt to consider the correlation between the activity and conformer, the accessible surface area (ASA) was calculated for each derivative to estimate the size or bulkiness of its conformation. Although the ASAs of nmr structures were not directly related to the type of conformer (figure 8 or square form), it was an important probe to consider the cytotoxicity of each derivative.
Doi
M
M
Osaka University of Pharmaceutical Sciences, Japan.
Shinozaki
F
F
In
Y
Y
Ishida
T
T
Yamamoto
D
D
Kamigauchi
M
M
Sugiura
M
M
Hamada
Y
Y
Kohda
K
K
Shioiri
T
T
D019943:Amino Acid Substitution; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D018360:Crystallography, X-Ray; D003603:Cytotoxins / Q000737:chemistry* / Q000633:toxicity; D007532:Isoleucine / Q000737:chemistry; D007939:Leukemia L1210; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000633:toxicity; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013816:Thermodynamics
D003603:Cytotoxins; D010456:Peptides, Cyclic; D007532:Isoleucine; C075813:ascidiacyclamide