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Updated: Jul 7, 2026

Molecular Spring Constant Analysis by Biomembrane Force Probe Spectroscopy
Published on: November 20, 2021
Demonstration by burst-phase analysis of a robust folding intermediate in the FF domain
Per Jemth1, Christopher M Johnson, Stefano Gianni
1MRC Centre for Protein Engineering, Hills Road, Cambridge CB2 2QH, UK.
Abstract:
The role of intermediates in the folding reaction of single-domain proteins is a controversial issue. It was previously shown by different methods that an on-pathway intermediate is populated in the presence of sodium sulphate during the folding of the FF domain from HYPA/FBP11. Here we demonstrate using analysis of the amplitudes of kinetic traces that this burst-phase folding intermediate is present at different salt concentration and at various pH, and is also found in roughly 30 site-directed mutants. The intermediate appears robust to changing conditions and thus fulfils an important criterion for a productive molecular species on the folding reaction pathway.
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