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© Research
Publication : Journal of biomolecular structure & dynamics

Reassessment of structural characteristics of the d(CGCG)2:actinomycin D complex from complete 1H and 31P NMR

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Journal of biomolecular structure & dynamics - 01 Dec 1989

Delepierre M, Van Heijenoort C, Igolen J, Pothier J, Le Bret M, Roques BP

Link to Pubmed [PMID] – 2627299

J. Biomol. Struct. Dyn. 1989 Dec;7(3):557-89

Complexes formed between Actinomycin D (ActD) and the tetranucleotides d(AGCT)2 and d(CGCG)2 were studied in detail by one and two-dimensional 1H and 31P NMR. The 31P two dimensional chemical exchange experiment, at room temperature on saturated complexes (1:1), showed unambiguously that the asymmetrical phenoxazone ring binds to the unique GC site under the two possible orientations in the d(AGCT)2 tetranucleotide but adopts a single orientation in the d(CGCG)2 tetranucleotide. For the d(CGCG)2:Act D saturated complex, complete assignments of all protons and phosphorus signals of the two-nucleotide strands, as well as of the two cyclic pentapeptide chains has allowed us to study in details the conformational features of the complex from NOE and coupling constants analysis. The tetranucleotide remains in a right-handed duplex, but the sugar puckers are modified for residues at the intercalation site. A uniform C2′ endo pucker is observed for residues on the strand facing the quinoid side of the phenoxazone ring while a C2′ endo-C3-endo equilibrium about 60% of C2′ endo is proposed for the two residues on the strand facing the benzenoid side of the phenoxazone ring. In contrast to previous studies on ActD-DNA interactions, we have been able to measure the 3J phosphorus-proton coupling constants at the intercalation site but also adjacent to it, showing that 31P chemical shifts are not simply related to the backbone conformation. Molecular mechanics calculations, using empirical distances deduced from NOE effects as restrained distances during minimizations, led to a model differing mainly from those previously published by orientation of the N methyl groups of both N-Methyl-Valines.

http://www.ncbi.nlm.nih.gov/pubmed/2627299