Protocol for the thermodynamic analysis of some proteins using an H/D exchange- and mass spectrometry-based technique.

Journal Article

SUPREX (stability of unpurified proteins from rates of H/D exchange) is a new H/D exchange- and mass spectrometry-based technique for the measurement of protein folding free energies (i.e., DeltaG values) and protein folding m values (i.e., deltaDeltaG/delta[denaturant]). Robust protocols for the acquisition and analysis of SUPREX data have been established and shown to be useful for the analysis of a number of different protein systems. Here we report on the SUPREX behavior of a special class of proteins that are not amenable to conventional SUPREX analyses using previously established protocols. This class of proteins includes protein systems that require an extended time to reach a folding/unfolding equilibrium in chemical denaturant-induced equilibrium unfolding experiments. As part of this work we use ubiquitin as a model system to highlight the complications that can arise in the conventional SUPREX analysis of such protein systems, and we describe a modified SUPREX protocol that can be used to eliminate these complications.

Full Text

Duke Authors

Cited Authors

  • Dai, SY; Gardner, MW; Fitzgerald, MC

Published Date

  • January 15, 2005

Published In

Volume / Issue

  • 77 / 2

Start / End Page

  • 693 - 697

PubMed ID

  • 15649073

International Standard Serial Number (ISSN)

  • 0003-2700

Digital Object Identifier (DOI)

  • 10.1021/ac048967z

Language

  • eng

Conference Location

  • United States