Accuracy and precision of a new H/D exchange- and mass spectrometry-based technique for measuring the thermodynamic properties of protein-peptide complexes.
A new H/D exchange- and MALDI mass spectrometry-based technique, termed SUPREX, was used to characterize the thermodynamic properties of a series of model protein-peptide complexes of the Abelson tyrosine kinase SH3 domain (abl-SH3) and the S-Protein (S-Pro). The SUPREX technique was employed to evaluate the folding free energies (DeltaG(f) values) of each model protein in the absence and in the presence of a series of different peptide ligands. Ultimately, these SUPREX-derived DeltaG(f) values were used to calculate dissociation constants (K(d) values) for each of the nine protein-peptide complexes in this study. As part of this work, we describe a new data collection and analysis method that allows the accurate and precise determination of protein folding m-values in the SUPREX experiment. The m-values that we determined for the abl-SH3 domain and the S-Pro system were in good agreement with those determined by conventional techniques. Our results also indicate that the SUPREX-derived K(d) values for the protein-peptide complexes in this work were in reasonably good agreement with those determined by conventional techniques.
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Related Subject Headings
- src Homology Domains
- Urea
- Thermodynamics
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
- Ribonucleases
- Ribonuclease, Pancreatic
- Peptide Fragments
- Molecular Sequence Data
- Ligands
- Kinetics
Citation
Published In
DOI
EISSN
ISSN
Publication Date
Volume
Issue
Start / End Page
Related Subject Headings
- src Homology Domains
- Urea
- Thermodynamics
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
- Ribonucleases
- Ribonuclease, Pancreatic
- Peptide Fragments
- Molecular Sequence Data
- Ligands
- Kinetics