Mutatational analysis of protein folding mechanisms

Abstract

Site-directed mutagenesis has proved to be a powerful technique for probing the relationship between the amino acid sequence of a protein and its folding, stability, structure, and function. This chapter presents the mutational analysis of protein folding mechanisms. The purpose is (1) to extend the approach to a three-state folding model involving a transient intermediate; (2) to demonstrate the way the integration of kinetic and equilibrium folding data can provide a comprehensive understanding of the effects of mutations on the folding mechanism; and (3) to summarize some of the recent findings on the folding of dihydrofolate reductase using this approach. The free energy data from both equilibrium and kinetic studies of the folding reactions of mutant and wild type proteins can be summarized in a reaction coordinate diagram. In this diagram, the free energies of the various species along the folding pathway are plotted as a function of the reaction coordinate. This coordinate does not correspond to any particular molecular dimension but rather reflects the progress of the folding reaction between the two end states, the native and the unfolded conformations.

Published in

Methods in Enzymology
1991, volume: 202, pages: 113-126

SLU Authors

• Finn, Bryan

  • Lund University

UKÄ Subject classification

Biophysics

Publication identifier

• DOI: https://doi.org/10.1016/0076-6879(91)02009-X

Permanent link to this page (URI)

https://res.slu.se/id/publ/132603