Studying biomolecular folding and binding using temperature-jump mass spectrometry.

Area of Science:

• Biochemistry
• Biophysics
• Analytical Chemistry

Background:

• Understanding biomolecular folding and complex formation is crucial for elucidating thermodynamics, kinetics, and folding pathways.
• Kinetic intermediates are significant as they represent potential drug targets.
• Native mass spectrometry offers a unique advantage by identifying and quantifying coexisting species, unlike conventional methods.

Purpose of the Study:

• To design and validate a temperature-jump electrospray source for mass spectrometry.
• To enable fast kinetics experiments for studying biomolecular folding and unfolding.
• To investigate the thermodynamics and kinetics of various biological systems, including peptides, proteins, and DNA complexes.

Main Methods:

• Development of a temperature-jump electrospray source for mass spectrometry.
• Performing fast kinetics experiments (0.16–32 s) across a temperature range of 10–90 °C.
• Utilizing temperature jumps (high-to-low and low-to-high) to record folding and unfolding kinetics.
• Applying the device to six biological systems, including peptides, proteins, and DNA complexes.

Main Results:

• The developed temperature-jump electrospray source successfully enables fast kinetics experiments.
• The setup allows for the recording of both folding and unfolding kinetics.
• Detailed thermodynamic and kinetic information was obtained for the folding and unfolding of a DNA triplex.

Conclusions:

• The novel temperature-jump electrospray source is a powerful tool for studying biomolecular kinetics.
• This method provides valuable insights into the thermodynamics and folding pathways of diverse biological molecules.
• The device facilitates the characterization of kinetic intermediates, potentially aiding drug discovery efforts.