This title appears in the Scientific Report :
2019
Please use the identifier:
http://hdl.handle.net/2128/24243 in citations.
Please use the identifier: http://dx.doi.org/10.1042/BCJ20190364 in citations.
Biochemical and structural characterization of murine GBP7, a guanylate binding protein with an elongated C-terminal tail
Biochemical and structural characterization of murine GBP7, a guanylate binding protein with an elongated C-terminal tail
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 b...
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Personal Name(s): | Legewie, Larissa |
---|---|
Loschwitz, Jennifer / Steffens, Nora / Prescher, Martin / Wang, Xue / Smits, Sander H. J. / Schmitt, Lutz / Strodel, Birgit / Degrandi, Daniel (Corresponding author) / Pfeffer, Klaus (Corresponding author) | |
Contributing Institute: |
JARA - HPC; JARA-HPC Strukturbiochemie; ICS-6 |
Published in: | Biochemical journal, 476 (2019) 21, S. 3161 - 3182 |
Imprint: |
London
Portland Press79505
2019
|
DOI: |
10.1042/BCJ20190364 |
PubMed ID: |
31689351 |
Document Type: |
Journal Article |
Research Program: |
Structural dynamics of murine guanylate binding proteins, their dimerization and interaction with lipid bilayers Functional Macromolecules and Complexes |
Link: |
Published on 2019-11-05. Available in OpenAccess from 2020-11-05. Published on 2019-11-05. Available in OpenAccess from 2020-11-05. |
Publikationsportal JuSER |
Please use the identifier: http://dx.doi.org/10.1042/BCJ20190364 in citations.
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. |