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Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics
Published on: April 17, 2017
Probing nucleic acid-ion interactions with buffer exchange-atomic emission spectroscopy.
Max Greenfeld1, Daniel Herschlag
1Department of Chemical Engineering and Biochemistry, Stanford University, Stanford, California, USA.
Buffer exchange-atomic emission spectroscopy (BE-AES) offers a new way to study nucleic acid ion atmospheres. This technique accurately measures all ions, overcoming limitations of older methods for biochemical and biophysical studies.
Area of Science:
- Biochemistry
- Biophysics
- Analytical Chemistry
Background:
- The ion atmosphere surrounding nucleic acids significantly influences their biochemical and biophysical characteristics.
- Studying this ion atmosphere is challenging due to its inherent diffuse and dynamic nature.
- Existing analytical techniques possess limitations in sensitivity, specificity, and directness for assaying ion atmospheres.
Purpose of the Study:
- To introduce Buffer Exchange-Atomic Emission Spectroscopy (BE-AES) as a novel technique for analyzing nucleic acid ion atmospheres.
- To overcome the limitations of conventional methods in quantifying ions associated with nucleic acids.
- To demonstrate the capability of BE-AES in providing a comprehensive analysis of ionic environments.
Main Methods:
- Development and application of Buffer Exchange-Atomic Emission Spectroscopy (BE-AES).
- Utilizing BE-AES to achieve a complete accounting of all ions within the ionic atmosphere of nucleic acids.
- Applying the technique to study ions at thermodynamic equilibrium.
Main Results:
- BE-AES provides a complete quantification of ions in the nucleic acid ionic atmosphere.
- The technique surpasses the sensitivity, specificity, and directness limitations of prior methods.
- Demonstrated successful application beyond nucleic acids to site-bound ions in RNA and proteins.
Conclusions:
- BE-AES is a powerful and versatile technique for the detailed study of ion atmospheres.
- This method enhances the understanding of nucleic acid properties by accurately characterizing their ionic environment.
- The applicability of BE-AES extends to studying bound ions in other biomolecules like RNA and proteins.

