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 Q. Hoang, Michael R. Baldwin, Karen R. Khar, John Karanicolas, Mark T. Fisher

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages293-304
Number of pages12
Volume1873
DOIs
StatePublished - 2019

Publication series

NameMethods in molecular biology (Clifton, N.J.)
PublisherHumana Press Inc.
ISSN (Print)1064-3745

Keywords

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

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