An optics-based variable-temperature assay system for characterizing thermodynamics of biomolecular reactions on...

Area of Science:

• Biochemistry
• Biophysics
• Chemical Thermodynamics

Background:

• Biological systems rely on the equilibrium of numerous biomolecular reactions.
• Physiological temperature (10-50 °C) significantly influences the importance and kinetics of these reactions.
• Understanding reaction thermodynamics provides insights into biomolecular processes.

Purpose of the Study:

• To develop a system for real-time measurement of multiple biomolecular binding reactions over a variable temperature range.
• To investigate the thermodynamics of plant lectin-glycan interactions.
• To elucidate the role of water molecules in the thermodynamics of these binding reactions.

Main Methods:

• Development of a variable-temperature opto-fluidic system capable of precise temperature control (±0.1 °C).
• Real-time measurement of 400-10,000 biomolecular binding reactions on solid supports.
• System validation using plant lectins and 24 synthetic glycans across temperatures from 10 °C to 60 °C.

Main Results:

• The developed system enables precise, real-time measurement of numerous binding reactions over an extended temperature range.
• Plant lectin-glycan binding reactions were characterized thermodynamically.
• Results indicated that these reactions can be enthalpy-driven, entropy-driven, or a combination of both, with significant influence from water molecules.

Conclusions:

• The variable-temperature opto-fluidic system is effective for characterizing the thermodynamics of biomolecular binding reactions.
• Lectin-glycan interactions exhibit diverse thermodynamic profiles.
• Water molecules are critical determinants of the thermodynamic landscape in lectin-glycan binding.