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@ARTICLE{Schulte:845271,
author = {Schulte, Marianne and Petrović, Dušan and Neudecker,
Philipp and Hartmann, Rudolf and Pietruszka, Jörg and
Willbold, Sabine and Willbold, Dieter and Panwalkar, Vineet},
title = {{C}onformational {S}ampling of the {I}ntrinsically
{D}isordered {C}-{T}erminal {T}ail of {DERA} {I}s
{I}mportant for {E}nzyme {C}atalysis},
journal = {ACS catalysis},
volume = {8},
number = {5},
issn = {2155-5435},
address = {Washington, DC},
publisher = {ACS},
reportid = {FZJ-2018-02552},
pages = {3971 - 3984},
year = {2018},
abstract = {2-Deoxyribose-5-phosphate aldolase (DERA) catalyzes the
reversible conversion of acetaldehyde and
glyceraldehyde-3-phosphate into deoxyribose-5-phosphate.
DERA is used as a biocatalyst for the synthesis of drugs
such as statins and is a promising pharmaceutical target due
to its involvement in nucleotide catabolism. Despite
previous biochemical studies suggesting the catalytic
importance of the C-terminal tyrosine residue found in
several bacterial DERAs, the structural and functional basis
of its participation in catalysis remains elusive because
the electron density for the last eight to nine residues
(i.e., the C-terminal tail) is absent in all available
crystal structures. Using a combination of NMR spectroscopy
and molecular dynamics simulations, we conclusively show
that the rarely studied C-terminal tail of E. coli DERA
(ecDERA) is intrinsically disordered and exists in
equilibrium between open and catalytically relevant closed
states, where the C-terminal tyrosine (Y259) enters the
active site. Nuclear Overhauser effect distance restraints,
obtained due to the presence of a substantial closed state
population, were used to derive the solution-state structure
of the ecDERA closed state. Real-time NMR hydrogen/deuterium
exchange experiments reveal that Y259 is required for
efficiency of the proton abstraction step of the catalytic
reaction. Phosphate titration experiments show that, in
addition to the phosphate-binding residues located near the
active site, as observed in the available crystal
structures, ecDERA contains previously unknown auxiliary
phosphate-binding residues on the C-terminal tail which
could facilitate in orienting Y259 in an optimal position
for catalysis. Thus, we present significant insights into
the structural and mechanistic importance of the ecDERA
C-terminal tail and illustrate the role of conformational
sampling in enzyme catalysis.},
cin = {ICS-6 / IBG-1 / IBOC / ZEA-3 / JARA-HPC},
ddc = {540},
cid = {I:(DE-Juel1)ICS-6-20110106 / I:(DE-Juel1)IBG-1-20101118 /
I:(DE-Juel1)IBOC-20090406 / I:(DE-Juel1)ZEA-3-20090406 /
$I:(DE-82)080012_20140620$},
pnm = {551 - Functional Macromolecules and Complexes (POF3-551) /
Computational Enzyme Design $(jics69_20151101)$},
pid = {G:(DE-HGF)POF3-551 / $G:(DE-Juel1)jics69_20151101$},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000431727300029},
pubmed = {pmid:30101036},
doi = {10.1021/acscatal.7b04408},
url = {https://juser.fz-juelich.de/record/845271},
}