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@ARTICLE{Cerminara:873619,
author = {Cerminara, Michele and Schöne, Antonie and Ritter, Ilona
and Gabba, Matteo and Fitter, Jörg},
title = {{M}apping {M}ultiple {D}istances in a {M}ultidomain
{P}rotein for the {I}dentification of {F}olding
{I}ntermediates},
journal = {Biophysical journal},
volume = {118},
number = {3},
issn = {0006-3495},
address = {Bethesda, Md.},
publisher = {Soc.},
reportid = {FZJ-2020-00862},
pages = {688-697},
year = {2020},
abstract = {The investigation and understanding of the folding
mechanism of multidomain proteins is still a challenge in
structural biology. The use of single-molecule Förster
resonance energy transfer offers a unique tool to map
conformational changes within the protein structure. Here,
we present a study following denaturant-induced unfolding
transitions of yeast phosphoglycerate kinase by mapping
several inter- and intradomain distances of this two-domain
protein, exhibiting a quite heterogeneous behavior. On the
one hand, the development of the interdomain distance during
the unfolding transition suggests a classical two-state
unfolding behavior. On the other hand, the behavior of some
intradomain distances indicates the formation of a compact
and transient molten globule intermediate state.
Furthermore, different intradomain distances measured within
the same domain show pronounced differences in their
unfolding behavior, underlining the fact that the choice of
dye attachment positions within the polypeptide chain has a
substantial impact on which unfolding properties are
observed by single-molecule Förster resonance energy
transfer measurements. Our results suggest that, to fully
characterize the complex folding and unfolding mechanism of
multidomain proteins, it is necessary to monitor multiple
intra- and interdomain distances because a single reporter
can lead to a misleading, partial, or oversimplified
interpretation.},
cin = {ICS-5},
ddc = {570},
cid = {I:(DE-Juel1)ICS-5-20110106},
pnm = {551 - Functional Macromolecules and Complexes (POF3-551)},
pid = {G:(DE-HGF)POF3-551},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:31916943},
UT = {WOS:000511291400016},
doi = {10.1016/j.bpj.2019.12.006},
url = {https://juser.fz-juelich.de/record/873619},
}