A similar in vitro and in cell lysate folding intermediate for the FF domain

Michael P. Latham, Lewis E. Kay

Research output: Contribution to journalArticlepeer-review

13 Scopus citations


Understanding the mechanisms by which proteins fold into their three-dimensional structures, including a description of the intermediates that are formed during the folding process, remains a goal of protein science. Most studies are performed under carefully controlled conditions in which the folding reaction is monitored in a buffer solution that is far from the natural milieu of the cell. Here, we have used (13)C and (1)H relaxation dispersion NMR spectroscopy to study folding of the FF domain in both Escherichia coli and Saccharomyces cerevisiae cellular lysates. We find that a conformationally excited state is populated in both lysates, which is very similar in structure to a folding intermediate observed in previous studies in buffer, with the kinetics and thermodynamics of the interconversion between native and intermediate conformers somewhat changed. The results point to the importance of extending folding studies beyond the test tube yet emphasize that insights can be obtained through careful experiments recorded in controlled buffer solutions.

Original languageEnglish (US)
Pages (from-to)3214-3220
Number of pages7
JournalJournal of Molecular Biology
Issue number19
StatePublished - Sep 23 2014
Externally publishedYes

Bibliographical note

Publisher Copyright:
Copyright © 2014 Elsevier Ltd. All rights reserved.


  • CPMG relaxation dispersion
  • in cell-like NMR
  • methyl side-chain
  • protein folding


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