Three steps in the thermal unfolding of F-actin: an experimental evidence.

Abstract:

:The development of differential scanning calorimetry has resulted in an increased interest in studies of the unfolding process in proteins with the aim of identifying domains and interactions with ligands or other proteins. Several of these studies were done with actin and showed that the thermal unfolding of F-actin occurs in at least three steps; this was interpreted as the denaturation of independent domains. In the present work, we have followed the thermal unfolding of F-actin using differential scanning calorimetry (DSC), CD spectroscopy, and probe fluorescence. We found that the three steps revealed through DSC are not the denaturation of independent domains. These three steps are a change in the environment of cys 374 at 49.5 degrees C; a modification at the nucleotide-binding site at 55 degrees C; and the unfolding of the peptide chain at 64 degrees C. Previous interpretations of the thermograms of F-actin were thus erroneous. Since DSC is now widely used to study proteins, our experimental approach and conclusions may also be relevant in denaturation studies of proteins in general.

journal_name

Biopolymers

journal_title

Biopolymers

authors

Gicquaud CR,Heppell B

doi

10.1002/bip.20567

subject

Has Abstract

pub_date

2006-11-01 00:00:00

pages

374-80

issue

4

eissn

0006-3525

issn

1097-0282

journal_volume

83

pub_type

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