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@ARTICLE{Lunev:421070,
author = {Lunev, Sergey and Butzloff, Sabine and Romero, Atilio R.
and Linzke, Marleen and Batista, Fernando A. and Meissner,
Kamila A. and Müller, Ingrid B. and Adawy, Alaa and
Wrenger, Carsten and Groves, Matthew R.},
title = {{O}ligomeric interfaces as a tool in drug discovery:
{S}pecific interference with activity of malate
dehydrogenase of {P}lasmodium falciparum in vitro},
journal = {PLOS ONE},
volume = {13},
number = {4},
issn = {1932-6203},
address = {San Francisco, California, US},
publisher = {PLOS},
reportid = {PUBDB-2019-01835},
pages = {e0195011 -},
year = {2018},
abstract = {Malaria remains a major threat to human health, as strains
resistant to current therapeutics are discovered. Efforts in
finding new drug targets are hampered by the lack of
sufficiently specific tools to provide target validation
prior to initiating expensive drug discovery projects. Thus,
new approaches that can rapidly enable drug target
validation are of significant interest.In this manuscript we
present the crystal structure of malate dehydrogenase from
Plasmodium falciparum (PfMDH) at 2.4 Å resolution and
structure-based mutagenic experiments interfering with the
inter-oligomeric interactions of the enzyme. We report
decreased thermal stability, significantly decreased
specific activity and kinetic parameters of PfMDH mutants
upon mutagenic disruption of either oligomeric interface. In
contrast, stabilization of one of the interfaces resulted in
increased thermal stability, increased substrate/cofactor
affinity and hyperactivity of the enzyme towards malate
production at sub-millimolar substrate concentrations.
Furthermore, the presented data show that our designed PfMDH
mutant could be used as specific inhibitor of the wild type
PfMDH activity, as mutated PfMDH copies were shown to be
able to self-incorporate into the native assembly upon
introduction in vitro, yielding deactivated mutant:wild-type
species. These data provide an insight into the role of
oligomeric assembly in regulation of PfMDH activity and
reveal that recombinant mutants could be used as probe tool
for specific modification of the wild type PfMDH activity,
thus offering the potential to validate its druggability in
vivo without recourse to complex genetics or initial tool
compounds. Such tool compounds often lack specificity
between host or pathogen proteins (or are toxic in in vivo
trials) and result in difficulties in assessing cause and
effect—particularly in cases when the enzymes of interest
possess close homologs within the human host. Furthermore,
our oligomeric interference approach could be used in the
future in order to assess druggability of other challenging
human pathogen drug targets.},
cin = {EMBL-User},
ddc = {610},
cid = {I:(DE-H253)EMBL-User-20120814},
pnm = {899 - ohne Topic (POF3-899)},
pid = {G:(DE-HGF)POF3-899},
experiment = {EXP:(DE-H253)D-K1.1-20150101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:29694407},
UT = {WOS:000430802400020},
doi = {10.1371/journal.pone.0195011},
url = {https://bib-pubdb1.desy.de/record/421070},
}