Researchers led by Dr. Fangjun Wang at the Dalian Institute of Chemical Physics, Chinese Academy of Sciences, have developed a time-resolved mass spectrometry method integrated with ultraviolet laser dissociation. This advancement enables the characterization of the stability and dynamic fine structure of target proteins caused by amino acid site mutations, offering a new technique for studying the pathological mechanisms of target protein amino acid mutations. The related findings have been published in the Journal of the American Chemical Society.
Understanding how amino acid site mutations affect the stability and dynamic structure of target proteins is crucial for unraveling the molecular mechanisms of diseases and designing targeted drugs. However, site mutations often only induce subtle changes in the dynamic structure of target proteins, posing significant challenges for characterizing their dynamic molecular details.
The research team employed an online mixing formic acid solution to induce the unfolding of target protein structures. Through non-denaturing electrospray ionization and mass spectrometric analysis, they monitored the charge distribution of protein unfolding intermediates and products, as well as their relative abundances with mixing time. Additionally, the team further utilized ultraviolet laser dissociation and fragment ion mass spectrometry quantification methods to compare the dynamic structures and molecular details of unfolding intermediates between mutant and wild-type dihydrofolate reductases.
This work provides a new technique for studying the subtle dynamic structures and pathological mechanisms of target proteins caused by amino acid site mutations.
Related Paper: https://doi.org/10.1021/jacs.4c00316
