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Thermodynamic stability measurements on multimeric proteins using a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-based method.

Publication ,  Journal Article
Powell, KD; Wales, TE; Fitzgerald, MC
Published in: Protein science : a publication of the Protein Society
April 2002

We recently reported on a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-based technique, termed SUPREX, that removes several important limitations associated with measuring the thermodynamic stability of proteins. In contrast to conventional spectroscopy-based techniques for characterizing the equilibrium unfolding behavior of proteins, SUPREX is amenable to the thermodynamic analysis of both purified and unpurified proteins using mg to ng quantities of material. Here we report on the application of SUPREX to the analysis of multimeric protein systems. Included in this work are the SUPREX results we obtained in studies on six model multimeric proteins including the GCN4p1 dimer, the coil-V(a)L(d) trimer, the 4-oxalocrotonate tautomerase (4-OT) hexamer, the Trp repressor (TrpR) dimer, the Arc repressor (ArcR) dimer, and an ArcR mutant (the (DOA20)ArcR) dimer which contained two destabilizing mutations including an Asp to Ala mutation at position 20 and an amide to ester bond mutation between amino acid (aa) residues 19 and 20. As part of the work described here, we present a new method for the analysis of SUPREX data that is generally applicable to both monomeric and multimeric protein systems. Our results on the model proteins in this study indicate that this new method can be used to determine folding free energies for proteins with the accuracy and precision of conventional spectroscopy-based methods.

Duke Scholars

Published In

Protein science : a publication of the Protein Society

DOI

EISSN

1469-896X

ISSN

0961-8368

Publication Date

April 2002

Volume

11

Issue

4

Start / End Page

841 / 851

Related Subject Headings

  • Viral Regulatory and Accessory Proteins
  • Viral Proteins
  • Thermodynamics
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Sensitivity and Specificity
  • Saccharomyces cerevisiae Proteins
  • Repressor Proteins
  • Protein Kinases
  • Protein Folding
  • Protein Conformation
 

Citation

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Powell, K. D., Wales, T. E., & Fitzgerald, M. C. (2002). Thermodynamic stability measurements on multimeric proteins using a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-based method. Protein Science : A Publication of the Protein Society, 11(4), 841–851. https://doi.org/10.1110/ps.3820102
Powell, Kendall D., Thomas E. Wales, and Michael C. Fitzgerald. “Thermodynamic stability measurements on multimeric proteins using a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-based method.Protein Science : A Publication of the Protein Society 11, no. 4 (April 2002): 841–51. https://doi.org/10.1110/ps.3820102.
Powell KD, Wales TE, Fitzgerald MC. Thermodynamic stability measurements on multimeric proteins using a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-based method. Protein science : a publication of the Protein Society. 2002 Apr;11(4):841–51.
Powell, Kendall D., et al. “Thermodynamic stability measurements on multimeric proteins using a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-based method.Protein Science : A Publication of the Protein Society, vol. 11, no. 4, Apr. 2002, pp. 841–51. Epmc, doi:10.1110/ps.3820102.
Powell KD, Wales TE, Fitzgerald MC. Thermodynamic stability measurements on multimeric proteins using a new H/D exchange- and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-based method. Protein science : a publication of the Protein Society. 2002 Apr;11(4):841–851.
Journal cover image

Published In

Protein science : a publication of the Protein Society

DOI

EISSN

1469-896X

ISSN

0961-8368

Publication Date

April 2002

Volume

11

Issue

4

Start / End Page

841 / 851

Related Subject Headings

  • Viral Regulatory and Accessory Proteins
  • Viral Proteins
  • Thermodynamics
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Sensitivity and Specificity
  • Saccharomyces cerevisiae Proteins
  • Repressor Proteins
  • Protein Kinases
  • Protein Folding
  • Protein Conformation