Imino proton exchange and base-pair kinetics in the AMP-RNA aptamer complex Journal Article
| Authors: | Nonin, S.; Jiang, F.; Patel, D. J. |
| Article Title: | Imino proton exchange and base-pair kinetics in the AMP-RNA aptamer complex |
| Abstract: | We report on the dynamics of base-pair opening in the ATP-binding asymmetric internal loop and flanking base-pairs of the AMP-RNA aptamer complex by monitoring the exchange characteristics of the extremely well resolved imino protons in the NMR spectrum of the complex. The kinetics of imino proton exchange as a function of basic pH or added ammonia catalyst are used to measure the apparent base-pair dissociation constants and lifetimes of Watson-Crick and mismatched base-pairs, as well as the solvent accessibility of the unpaired imino protons in the complex. The exchange characteristics of the imino protons identify the existence of four additional hydrogen bonds stabilizing the conformation of the asymmetric ATP-binding internal loop that were not detected by NOEs and coupling constants alone, but are readily accommodated in the previously reported solution structure of the AMP-RNA aptamer complex published from our laboratory. The hydrogen exchange kinetics of the non-Watson-Crick pairs in the asymmetric internal loop of the AMP-RNA aptamer complex have been characterized and yield apparent dissociation constants (αK(d)) that range from 10-2 to 10-7. Surprisingly, three of these αK(d), values are amongst the lowest measured for all base-pairs in the AMP-RNA aptamer complex. Comparative studies of hydrogen exchange of the imino protons in the free RNA aptamer and the AMP-RNA aptamer complex establish that complexation stabilizes not only the bases within the ATP-binding asymmetric internal loop, but also the flanking stem base-pairs (two pairs on either side) of the binding site. We also outline some preliminary results related to the exchange properties of a sugar 2'-hydroxyl proton of a guanosine residue involved in a novel hydrogen bond that has been shown to contribute to the immobilization of the bound AMP by the RNA aptamer, and whose resonance is narrow and downfield shifted in the spectrum. |
| Keywords: | nonhuman; rna; kinetics; nucleotide sequence; magnetic resonance spectroscopy; nuclear magnetic resonance spectroscopy; base pairing; hydrogen bond; hydrogen bonding; thermodynamics; catalysis; nucleic acid conformation; proton; adenosine phosphate; nucleic acid structure; dissociation constant; adenosine monophosphate; oligoribonucleotides; priority journal; article; imines; ion exchange; amp-rna aptamer complex; base-pair dissociation constants; imino proton exchange; mismatch pair opening; sugar 2'-oh groups; imino acid |
| Journal Title: | Journal of Molecular Biology |
| Volume: | 268 |
| Issue: | 2 |
| ISSN: | 0022-2836 |
| Publisher: | Academic Press Inc., Elsevier Science |
| Date Published: | 1997-05-02 |
| Start Page: | 359 |
| End Page: | 374 |
| Language: | English |
| DOI: | 10.1006/jmbi.1997.0986 |
| PUBMED: | 9159476 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 17 March 2017 -- Source: Scopus |
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