Integration and global analysis of isothermal titration calorimetry data for studying macromolecular interactions

Chad A Brautigam, Huaying Zhao, Carolyn Vargas, Sandro Keller, Peter Schuck

Research output: Contribution to journalArticle

84 Scopus citations

Abstract

Isothermal titration calorimetry (ITC) is a powerful and widely used method to measure the energetics of macromolecular interactions by recording a thermogram of differential heating power during a titration. However, traditional ITC analysis is limited by stochastic thermogram noise and by the limited information content of a single titration experiment. Here we present a protocol for bias-free thermogram integration based on automated shape analysis of the injection peaks, followed by combination of isotherms from different calorimetric titration experiments into a global analysis, statistical analysis of binding parameters and graphical presentation of the results. This is performed using the integrated public-domain software packages NITPIC, SEDPHAT and GUSSI. The recently developed low-noise thermogram integration approach and global analysis allow for more precise parameter estimates and more reliable quantification of multisite and multicomponent cooperative and competitive interactions. Titration experiments typically take 1-2.5 h each, and global analysis usually takes 10-20 min.

Original languageEnglish (US)
Pages (from-to)882-894
Number of pages13
JournalNature Protocols
Volume11
Issue number5
DOIs
StatePublished - May 1 2016

ASJC Scopus subject areas

  • Medicine(all)

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