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@ARTICLE{Legewie:873653,
author = {Legewie, Larissa and Loschwitz, Jennifer and Steffens, Nora
and Prescher, Martin and Wang, Xue and Smits, Sander H. J.
and Schmitt, Lutz and Strodel, Birgit and Degrandi, Daniel
and Pfeffer, Klaus},
title = {{B}iochemical and structural characterization of murine
{GBP}7, a guanylate binding protein with an elongated
{C}-terminal tail},
journal = {Biochemical journal},
volume = {476},
number = {21},
issn = {1470-8728},
address = {London},
publisher = {Portland Press79505},
reportid = {FZJ-2020-00884},
pages = {3161 - 3182},
year = {2019},
abstract = {Guanylate-binding proteins (GBPs) constitute a family of
interferon-inducible guanosine triphosphatases (GTPases)
that are key players in host defense against intracellular
pathogens ranging from protozoa to bacteria and viruses. So
far, human GBP1 and GBP5 as well as murine GBP2 (mGBP2) have
been biochemically characterized in detail. Here, with
murine GBP7 (mGBP7), a GBP family member with an
unconventional and elongated C-terminus is analyzed. The
present study demonstrates that mGBP7 exhibits a
concentration-dependent GTPase activity and an apparent GTP
turnover number of 20 min-1. In addition, fluorescence
spectroscopy analyses reveal that mGBP7 binds GTP with high
affinity (KD = 0.22 µM) and GTPase activity assays
indicate that mGBP7 hydrolyzes GTP to GDP and GMP. The mGBP7
GTPase activity is inhibited by incubation with γ-phosphate
analogs and a K51A mutation interfering with GTP binding.
SEC-MALS analyses give evidence that mGBP7 forms transient
dimers and that this oligomerization pattern is not
influenced by the presence of nucleotides. Moreover, a
structural model for mGBP7 is provided by homology modeling,
which shows that the GTPase possesses an elongated
C-terminal (CT) tail compared with the CaaX motif-containing
mGBP2 and human GBP1. Molecular dynamics simulations
indicate that this tail has transmembrane characteristics
and, interestingly, confocal microscopy analyses reveal that
the CT tail is required for recruitment of mGBP7 to the
parasitophorous vacuole of Toxoplasma gondii.},
cin = {ICS-6 / JARA-HPC},
ddc = {540},
cid = {I:(DE-Juel1)ICS-6-20110106 / $I:(DE-82)080012_20140620$},
pnm = {551 - Functional Macromolecules and Complexes (POF3-551) /
Structural dynamics of murine guanylate binding proteins,
their dimerization and interaction with lipid bilayers
$(jics6a_20190501)$},
pid = {G:(DE-HGF)POF3-551 / $G:(DE-Juel1)jics6a_20190501$},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:31689351},
UT = {WOS:000509877500004},
doi = {10.1042/BCJ20190364},
url = {https://juser.fz-juelich.de/record/873653},
}