Constructing Kinetically Controlled Denaturation Isotherms of Folded Proteins Using Denaturant-Pulse Chaperonin Binding

Methods Mol Biol. 2019:1873:293-304. doi: 10.1007/978-1-4939-8820-4_19.

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.

Keywords: Aggregation; Biolayer interferometry; Chaperonin; Denaturant pulse; Denaturation; GroEL; Protein folding.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Biosensing Techniques
  • Cell Line
  • Data Analysis
  • Kinetics
  • Protein Aggregates
  • Protein Binding
  • Protein Denaturation*
  • Protein Folding*
  • Proteins / chemistry*
  • Proteins / metabolism
  • Software
  • Temperature*
  • User-Computer Interface

Substances

  • Protein Aggregates
  • Proteins