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Updated: Aug 30, 2026

Monitoring the Reductive and Oxidative Half-Reactions of a Flavin-Dependent Monooxygenase using Stopped-Flow Spectrophotometry
Published on: March 18, 2012
Analysis of the stability of cytochrome c(6) with an improved stopped-flow protocol
Christian Lange1, Manuel Hervás, Miguel A De la Rosa
1Instituto de Bioquímica Vegetal y Fotosíntesis, Centro de Investigaciones Científicas Isla de la Cartuja, Universidad de Sevilla y CSIC, Avda. Américo Vespucio s/n, 41092 Sevilla, Spain.
Abstract:
This work presents an improved stopped-flow protocol for the simultaneous measurement of thermodynamic and kinetic protein stability data from a single experiment, along with a formalism for the global analysis of the data. The method was applied to the comparison of the stabilities of cytochrome c(6) from Anabaena sp. PCC 7119 and one of its mutants (D72K). Compared to the wild type the mutant was found to have a significantly reduced thermodynamic (deltadeltaG(U0)=2.7 kJ mol(-1)) and kinetic stability, as well as a more pronounced shift in transition state structure upon destabilization.

