Resolution of temperature dependent conformational multiequilibria processes

Multivariate Curve Resolution (MCR) is applied to the study of temperature dependent conformational multiequilibria evolving processes. Experimental data sets are obtained by UV spectral monitoring of the melting behavior of the heteropolynucleotide poly(adenylic) acid–poly(uridylic) acid (poly(A)–poly(U)) and of the hompolynucleotides poly(adenylic) (poly(A)) acid and poly(urydilic) acid (poly(U)), i.e., recording UV spectra at different temperatures during the melting process of these polynucleotides. Separate study of every individual melting experiment by MCR did not give a satisfactory resolution of the heteropolynucleotide poly(A)–poly(U) melting process because of unresolved rotational ambiguities and rank deficiency problems. Conversely, the simultaneous MCR analysis of the melting process of poly(A)–poly(U) heteropolynucleotide together with the separate melting processes of the poly(A) and poly(U) homopolynucleotides, allowed the resolution of the species profiles and the elimination of the rank deficiency problems present in the individual analysis of the melting behavior of poly(A)–poly(U).