Quantifying protein unfolding kinetics with a high-throughput microfluidic platform.

ABSTRACT: Even after folding, proteins transiently sample unfolded or partially unfolded intermediates, and these species are often at risk of irreversible alteration (e.g. via proteolysis, aggregation, or post-translational modification). Kinetic stability, in addition to thermodynamic stability, can directly impact protein lifetime, abundance, and the formation of alternative, sometimes disruptive states. However, we have very few measurements of protein unfolding rates or how mutations alter these rates, largely due to technical challenges associated with their measurement. To address this need, we developed SPARKfold (Simultaneous Proteolysis Assay Revealing Kinetics of Folding), a microfluidic platform to express, purify, and measure unfolding rate constants for >1000 protein variants in

SUBMITTER: Atsavapranee B 

PROVIDER: S-EPMC11761748 | biostudies-literature | 2025 Jan

REPOSITORIES: biostudies-literature