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2D-IR spectroscopy probes ions interacting with biomolecules

14 January 2019 | News

DNA and RNA are charged polymers with their negative charges located in the molecular backbone, which consists of ionic phosphate (PO2) and of sugar groups (Figure 1). Stabilisation of the macromolecular structures of DNA and RNA requires a compensation of strong repulsive electric forces between the equally charged phosphate groups by ions of opposite, i.e., positive charge. In this context, magnesium (Mg2+) ions are particularly relevant as they not only stabilise the structure, but also mediate the recognition of external binding partners and act as catalytic centres. Moreover, changes of macromolecular structure via dynamic folding processes are connected with a rearrangement of positive ions embedded in the surrounding water shell.

Positive ions are arranged in different geometries around DNA and RNA: in so-called site-bound or contact-pair geometries, a positive ion is located in direct contact with an oxygen atom of a phosphate group. In contrast, the so-called outer ion atmosphere consists of positive ions separated by at least one layer of water molecules from the phosphate groups. The functional role of the different geometries and the underlying interactions are far from being understood. A deeper insight at the molecular level requires highly sensitive probes which allow for discerning the different ion geometries without disturbing them, and for mapping their dynamics on the ultrafast time scale of molecular motions.

Figure 2In a recent publication in J. Phys. Chem. Letters, researchers from the Max Born Institute (MBI) demonstrate that vibrations of phosphate groups represent sensitive and non-invasive probes of ion geometries in a water environment. Dimethylphosphate [DMP, (CH3O)2PO2], an established model system for the DNA and RNA backbone, was prepared in liquid water with an excess of Mg2+ ions (Figure 2, top) and studied by non-linear vibrational spectroscopy in the femtosecond time domain. The experiments make use of two-dimensional infrared (2D-IR) spectroscopy.

The experiments map Mg2+ ions in direct contact with a PO2 group via a distinct feature in the 2D-IR spectrum (Figure 2, bottom). The interaction with the Mg2+ ion shifts the asymmetric PO2 stretching vibration to a frequency which is higher than in absence of Mg2+ ions. The lineshape and the time evolution of this new feature reveal fluctuations of the contact ion pair geometry and the embedding water shell on a time scale of hundreds of femtoseconds while the contact pair itself exists for much longer times (~10–6 s). An in-depth theoretical analysis shows that the subtle balance of attractive electrostatic (Coulomb) forces and repulsive forces due to the quantum-mechanical exchange interaction govern the frequency position of the phosphate vibration.

The ability of 2D-IR spectroscopy to characterise the short-ranged phosphate-ion interaction in solution provides a novel analytical tool that complements currently available structural techniques. An extension of this new approach to DNA and RNA and their ionic environment is most promising and expected to provide new insight in the forces stabilising equilibrium structures and driving folding processes.

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