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
Kendall D PowellMichael C Fitzgerald

Abstract

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 prote...Continue Reading

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Citations

Sep 5, 2002·Mass Spectrometry Reviews·Igor A Kaltashov, Stephen J Eyles
May 28, 2003·Mass Spectrometry Reviews·Lars Konermann, Douglas A Simmons
Mar 10, 2009·Journal of the American Society for Mass Spectrometry·Michael C Fitzgerald, Graham M West
May 7, 2010·Journal of the American Society for Mass Spectrometry·Tingting TuMichael L Gross
Apr 20, 2004·Current Opinion in Structural Biology·Jason Lanman, Peter E Prevelige
Apr 3, 2008·Nature Reviews. Drug Discovery·Malin C Lagerström, Helgi B Schiöth
Jun 5, 2014·Annual Review of Analytical Chemistry·M Ariel Geer, Michael C Fitzgerald
Mar 31, 2009·Analytical Biochemistry·John S Vrettos, Curtis W Meuse
Aug 24, 2004·Protein Science : a Publication of the Protein Society·Chiwook Park, Susan Marqusee
Jun 26, 2009·Protein Science : a Publication of the Protein Society·James A Mobley, Anton Poliakov
Mar 20, 2003·Journal of Mass Spectrometry : JMS·Marc Kipping, Angelika Schierhorn
Mar 15, 2008·Biochemistry·Petra L RoulhacMichael C Fitzgerald
Aug 29, 2002·Journal of the American Chemical Society·Kendall D PowellMichael C Fitzgerald
Mar 9, 2004·Journal of Combinatorial Chemistry·Kendall D Powell, Michael C Fitzgerald
Jul 31, 2004·Analytical Chemistry·Michael Z WangMichael C Fitzgerald
Dec 4, 2021·Mass Spectrometry Reviews·Annelaure DamontJean-Claude Tabet

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