Changes in solvent exposure reveal the kinetics and equilibria of adsorbed protein unfolding in hydrophobic interaction chromatography.

Journal of Chromatography. a
R W DeitcherErik J Fernandez

Abstract

Hydrogen exchange has been a useful technique for studying the conformational state of proteins, both in bulk solution and at interfaces, for several decades. Here, we propose a physically based model of simultaneous protein adsorption, unfolding and hydrogen exchange in HIC. An accompanying experimental protocol, utilizing mass spectrometry to quantify deuterium labeling, enables the determination of both the equilibrium partitioning between conformational states and pseudo-first order rate constants for folding and unfolding of adsorbed protein. Unlike chromatographic techniques, which rely on the interpretation of bulk phase behavior, this methodology utilizes the measurement of a molecular property (solvent exposure) and provides insight into the nature of the unfolded conformation in the adsorbed phase. Three model proteins of varying conformational stability, alpha-chymotrypsinogen A, beta-lactoglobulin B, and holo alpha-lactalbumin, are studied on Sepharose HIC resins possessing assorted ligand chemistries and densities. alpha-Chymotrypsinogen, conformationally the most stable protein in the set, exhibits no change in solvent exposure at all the conditions studied, even when isocratic pulse-response chromatography sugges...Continue Reading

References

Jan 1, 1991·The Biochemical Journal·S E RadfordC M Dobson
Nov 1, 1988·International Journal of Peptide and Protein Research·N K Kella, J E Kinsella
Apr 1, 1986·Journal of Biochemistry·M IkeguchiS Sugai
Jan 1, 1966·Advances in Protein Chemistry·A Hvidt, S O Nielsen
Nov 1, 1983·Quarterly Reviews of Biophysics·S W Englander, N R Kallenbach
Aug 1, 1984·Analytical Biochemistry·L Lindahl, H J Vogel
Jul 14, 1995·Science·Y BaiS W Englander
Apr 1, 1993·Protein Science : a Publication of the Protein Society·Z Zhang, D L Smith
Oct 1, 1995·Protein Science : a Publication of the Protein Society·J K MyersJ M Scholtz
Oct 1, 1996·International Journal of Biological Macromolecules·P AymardT Nicolai
Mar 26, 1998·International Journal of Biological Macromolecules·D RenardW G Griffin
May 30, 1998·Journal of Molecular Biology·K W PlaxcoD Baker
Mar 10, 1999·Biopolymers·M VerheulK G de Kruif
Nov 4, 2000·Biochimica Et Biophysica Acta·L Sawyer, G Kontopidis
Oct 23, 2001·Biotechnology and Bioengineering·J L McNayE J Fernandez
Jan 10, 2002·The Journal of Biological Chemistry·Maarten F M EngelAntonie J W G Visser
May 22, 2002·Protein Science : a Publication of the Protein Society·Scott A Tobler, Erik J Fernandez
Mar 26, 2003·Journal of Colloid and Interface Science·Tara Tibbs Jones, Erik J Fernandez
May 28, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Rainer HahnAlois Jungbauer
Jul 21, 2004·Proceedings of the National Academy of Sciences of the United States of America·Maarten F M EngelCarlo P M van Mierlo
Jul 26, 2005·Journal of Chromatography. a·Alois JungbauerRainer Hahn
Nov 23, 2006·Journal of Chromatography. a·Emmerich HaimerAlois Jungbauer
Aug 19, 2007·Journal of Chromatography. a·Jie Chen, Steven M Cramer
Aug 24, 2007·Proceedings of the National Academy of Sciences of the United States of America·Per ZetterströmStefan L Marklund
Jan 22, 2008·Bioprocess and Biosystems Engineering·Justin T McCueJörg Thömmes
Jun 11, 2008·Journal of Chromatography. a·Rene UeberbacherAlois Jungbauer
Aug 23, 2008·Journal of Chromatography. a·Brian C S To, Abraham M Lenhoff
Jun 9, 2009·Journal of Chromatography. a·Rene UeberbacherAlois Jungbauer

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Citations

Dec 3, 2014·Colloids and Surfaces. B, Biointerfaces·Patricia A SantosMauricio S Baptista
Oct 4, 2011·Journal of Chromatography. a·Patrícia P AguilarAna C Dias-Cabral
Nov 1, 2012·Journal of Separation Science·Rainer Hahn
Jan 21, 2015·Journal of Chromatography. a·Linling YuYan Sun
Dec 3, 2014·Journal of Chromatography. a·Adrian M GospodarekErik J Fernandez
Feb 9, 2019·Preparative Biochemistry & Biotechnology·Agnes RodlerAlois Jungbauer
Apr 11, 2018·Journal of Chromatography. a·Beate Beyer, Alois Jungbauer

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