Modern Kinetics and Mechanism of Protein Folding: A Retrospective

Modern experimental kinetics of protein folding began in the early 1990s with the introduction of nanosecond laser pulses to trigger the folding reaction, providing an almost 10(6)-fold improvement in time resolution over the stopped-flow method being employed at the time. These experiments marked the beginning of the fast-folding subfield that enabled investigation of the kinetics of formation of secondary structural elements and disordered loops for the first time, as well as the fastest folding proteins. When I started to work on this subject, a fast folding protein was one that folded in milliseconds. There were, moreover, no analytical theoretical models and no atomistic or coarse-grained molecular dynamics simulations to describe the mechanism. Two of the most important discoveries from my lab since then are a protein that folds in hundreds of nanoseconds, as determined from nanosecond laser temperature experiments, and the discovery that the theoretically predicted barrier crossing time is about the same for proteins that differ in folding rates by 10(4)-fold, as determined from single molecule fluorescence measurements. We also developed what has been called the Huckel model of protein folding, which quantitatively explains a wide range of equilibrium and kinetic measurements. This retrospective traces the history of contributions to the fast folding subfield from my lab until about 3 years ago, when I left protein folding to spend the rest of my research career trying to discover an inexpensive drug for treating sickle cell disease.

Automated summary

Modern experimental kinetics of protein folding were revolutionized in the early 1990s by the introduction of nanosecond laser pulses, leading to the investigation of fast-folding proteins and the dynamics of secondary structures. This development marked a significant improvement in time resolution and enabled the discovery of previously unknown folding mechanisms in protein kinetics research.