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10 pages, 4 figures, 1 scheme, 1 table.-- PMID: 12202780 [PubMed].-- PMCID: PMC137437.A successful application is reported of the multivariate curve resolution alternating least-squares method (MCR-ALS) for the analysis of nucleic acid melting and salt-induced transitions. Under conditions where several structures co-exist in a conformational equilibrium, MCR-ALS analysis of the UV and circular dichroism (CD) spectra at different temperatures, ionic strength and oligonucleotide concentration allows for the resolution of concentration profiles and pure spectra of the different species. The methodology is illustrated by the case of the cyclic oligonucleotide d<pTGCTCGCT>. The melting transition of this molecule at different oligonucleotide concentrations was studied at 0, 2 and 10 mM MgCl2 by UV and CD spectroscopy. In addition, salt titration experiments were carried out at 21.0 and 54.0°C. The MCR-ALS analysis indicates that three different conformations of this molecule co-exist in solution. In agreement with previous NMR studies, these conformations were assigned to a monomeric dumbbell-like structure, a dimeric four-stranded conformation and a disordered (random coil) structure. The MCR-ALS methodology allows for a detailed analysis of how this equilibrium is affected by temperature, salt and oligonucleotide concentration.This research was supported by the Spanish MCYT (BQU2000-0788 and BQU2001-3693) and the Generalitat de Catalunya (SGR-0048, SGR-0049, SGR-00056 and Centre de Referència de Biotecnologia).Peer reviewe

Multivariate curve resolution: a powerful tool for the analysis of conformational transitions in nucleic acids