Abstract

A new method for the determination of the kinetics of exchange of the imino protons of DNA duplex is reported using a combination NOESY and ROESY experiments at short mixing times (less-than-or-equal-to 20 ms). These results have been compared with the commonly used longitudinal relaxation approach through the T1 measurement. To calculate k(ex) and tau(ex) by ROESY-NOESY experiment, the volume of the cross-peaks between imino protons and water in the NOESY and ROESY spectra have been measured separately from the magnetization term. This work shows that the present approach for the measurement of the kinetics of slow exchanging imino protons of DNA duplex is comparable to the saturation recovery experiment in which the exchange rate can be accelerated by the addition of a base catalyst. The present ROESY-NOESY approach has been found to be particularly useful and reasonably accurate for the measurement of exchange kinetics of both the fast- and slow-exchanging imino protons in DNA duplex both under non-physiological and physiological condition where the saturation recovery method can not be used.

Published in

Nucleic Acids Research
1993, Volume: 21, number: 18, pages: 42954288
Publisher: OXFORD UNIV PRESS UNITED KINGDOM

UKÄ Subject classification

Structural Biology


Publication identifier

DOI: https://doi.org/10.1093/nar/21.18.4288

Permanent link to this page (URI)

https://res.slu.se/id/publ/83880