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@ARTICLE{Panwalkar:283504,
author = {Panwalkar, Vineet and Neudecker, Philipp and Schmitz,
Michael and Lecher, Justin and Schulte, Marianne and Medini,
Karima and Stoldt, Matthias and Brimble, Margaret A and
Willbold, Dieter and Dingley, Andrew J},
title = {{T}he {N}edd4-1 {WW} {D}omain {R}ecognizes the {PY} {M}otif
{P}eptide through {C}oupled {F}olding and {B}inding
{E}quilibria.},
journal = {Biochemistry},
volume = {55},
number = {4},
issn = {1520-4995},
address = {Columbus, Ohio},
publisher = {American Chemical Society},
reportid = {FZJ-2016-01837},
pages = {659 - 674},
year = {2016},
abstract = {The four WW domains of human Nedd4-1 (neuronal precursor
cell expressed developmentally downregulated gene 4-1)
interact with the PPxY (PY) motifs of the human epithelial
Na(+) channel (hENaC) subunits, with the third WW domain
(WW3*) showing the highest affinity. We have shown
previously that the α-hENaC PY motif binding interface of
WW3* undergoes conformational exchange on the millisecond
time scale, indicating that conformational sampling plays a
role in peptide recognition. To further understand this
role, the structure and dynamics of hNedd4-1 WW3* were
investigated. The nuclear Overhauser effect-derived
structure of apo-WW3* resembles the domain in complex with
the α-hENaC peptide, although particular side chain
conformations change upon peptide binding, which was further
investigated by molecular dynamics simulations. Model-free
analysis of the (15)N nuclear magnetic resonance spin
relaxation data showed that the apo and peptide-bound states
of WW3* have similar backbone picosecond to nanosecond time
scale dynamics. However, apo-WW3* exhibits pronounced
chemical exchange on the millisecond time scale that is
quenched upon peptide binding. (1)HN and (15)N
Carr-Purcell-Meiboom-Gill (CPMG) relaxation dispersion
experiments at various temperatures revealed that apo-WW3*
exists in an equilibrium between the natively folded peptide
binding-competent state and a random coil-like denatured
state. The thermodynamics of the folding equilibrium was
determined by fitting a thermal denaturation profile
monitored by circular dichroism spectroscopy in combination
with the CPMG data, leading to the conclusion that the
unfolded state is populated to $∼20\%$ at 37 °C. These
results show that the binding of the hNedd4-1 WW3* domain to
α-hENaC is coupled to the folding equilibrium.},
cin = {ICS-6},
ddc = {570},
cid = {I:(DE-Juel1)ICS-6-20110106},
pnm = {553 - Physical Basis of Diseases (POF3-553)},
pid = {G:(DE-HGF)POF3-553},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:26685112},
UT = {WOS:000369471800005},
doi = {10.1021/acs.biochem.5b01028},
url = {https://juser.fz-juelich.de/record/283504},
}