Constructing kinetically controlled denaturation isotherms of folded proteins using denaturant-pulse chaperonin binding

Pierce T. O’Neil, Alexandra J. Machen, Jackie A. Thompson, Wei Wang, Quyen Hoang, Michael R. Baldwin, Karen R. Khar, John Karanicolas, Mark T. Fisher

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Methods to assess the kinetic stability of proteins, particularly those that are aggregation prone, are very useful in establishing ligand induced stabilizing effects. Because aggregation prone proteins are by nature difficult to work with, most solution based methods are compromised by this inherent instability. Here, we describe a label-free method that examines the denaturation of immobilized proteins where the dynamic unfolded protein populations are captured and detected by chaperonin binding.

LanguageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages293-304
Number of pages12
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1873
ISSN (Print)1064-3745

Fingerprint

Chaperonins
Immobilized Proteins
Protein Unfolding
Proteins
Protein Stability
Ligands
Population

Keywords

  • Aggregation
  • Biolayer interferometry
  • Chaperonin
  • Denaturant pulse
  • Denaturation
  • GroEL
  • Protein folding

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

O’Neil, P. T., Machen, A. J., Thompson, J. A., Wang, W., Hoang, Q., Baldwin, M. R., ... Fisher, M. T. (2019). Constructing kinetically controlled denaturation isotherms of folded proteins using denaturant-pulse chaperonin binding. In Methods in Molecular Biology (pp. 293-304). (Methods in Molecular Biology; Vol. 1873). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8820-4_19

Constructing kinetically controlled denaturation isotherms of folded proteins using denaturant-pulse chaperonin binding. / O’Neil, Pierce T.; Machen, Alexandra J.; Thompson, Jackie A.; Wang, Wei; Hoang, Quyen; Baldwin, Michael R.; Khar, Karen R.; Karanicolas, John; Fisher, Mark T.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 293-304 (Methods in Molecular Biology; Vol. 1873).

Research output: Chapter in Book/Report/Conference proceedingChapter

O’Neil, PT, Machen, AJ, Thompson, JA, Wang, W, Hoang, Q, Baldwin, MR, Khar, KR, Karanicolas, J & Fisher, MT 2019, Constructing kinetically controlled denaturation isotherms of folded proteins using denaturant-pulse chaperonin binding. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1873, Humana Press Inc., pp. 293-304. https://doi.org/10.1007/978-1-4939-8820-4_19
O’Neil PT, Machen AJ, Thompson JA, Wang W, Hoang Q, Baldwin MR et al. Constructing kinetically controlled denaturation isotherms of folded proteins using denaturant-pulse chaperonin binding. In Methods in Molecular Biology. Humana Press Inc. 2019. p. 293-304. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-8820-4_19
O’Neil, Pierce T. ; Machen, Alexandra J. ; Thompson, Jackie A. ; Wang, Wei ; Hoang, Quyen ; Baldwin, Michael R. ; Khar, Karen R. ; Karanicolas, John ; Fisher, Mark T. / Constructing kinetically controlled denaturation isotherms of folded proteins using denaturant-pulse chaperonin binding. Methods in Molecular Biology. Humana Press Inc., 2019. pp. 293-304 (Methods in Molecular Biology).
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