Domain motions, dimerization, and membrane interactions of the murine guanylate binding protein 2
Abstract Guanylate-binding proteins (GBPs) are a group of GTPases that are induced by interferon- $$\gamma$$ γ and are crucial components of cell-autonomous immunity against intracellular pathogens. Here, we examine murine GBP2 (mGBP2), which we have previously shown to be an essential effector prot...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-01-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-27520-8 |
| _version_ | 1797952467773685760 |
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| author | Jennifer Loschwitz Nora Steffens Xue Wang Moritz Schäffler Klaus Pfeffer Daniel Degrandi Birgit Strodel |
| author_facet | Jennifer Loschwitz Nora Steffens Xue Wang Moritz Schäffler Klaus Pfeffer Daniel Degrandi Birgit Strodel |
| author_sort | Jennifer Loschwitz |
| collection | DOAJ |
| description | Abstract Guanylate-binding proteins (GBPs) are a group of GTPases that are induced by interferon- $$\gamma$$ γ and are crucial components of cell-autonomous immunity against intracellular pathogens. Here, we examine murine GBP2 (mGBP2), which we have previously shown to be an essential effector protein for the control of Toxoplasma gondii replication, with its recruitment through the membrane of the parasitophorous vacuole and its involvement in the destruction of this membrane likely playing a role. The overall aim of our work is to provide a molecular-level understanding of the mutual influences of mGBP2 and the parasitophorous vacuole membrane. To this end, we performed lipid-binding assays which revealed that mGBP2 has a particular affinity for cardiolipin. This observation was confirmed by fluorescence microscopy using giant unilamellar vesicles of different lipid compositions. To obtain an understanding of the protein dynamics and how this is affected by GTP binding, mGBP2 dimerization, and membrane binding, assuming that each of these steps are relevant for the function of the protein, we carried out standard as well as replica exchange molecular dynamics simulations with an accumulated simulation time of more than 30 μs. The main findings from these simulations are that mGBP2 features a large-scale hinge motion in its M/E domain, which is present in each of the studied protein states. When bound to a cardiolipin-containing membrane, this hinge motion is particularly pronounced, leading to an up and down motion of the M/E domain on the membrane, which did not occur on a membrane without cardiolipin. Our prognosis is that this up and down motion has the potential to destroy the membrane following the formation of supramolecular mGBP2 complexes on the membrane surface. |
| first_indexed | 2024-04-10T22:47:54Z |
| format | Article |
| id | doaj.art-2733771ef0b34728a4430cdf7c2f7000 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-10T22:47:54Z |
| publishDate | 2023-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-2733771ef0b34728a4430cdf7c2f70002023-01-15T12:12:58ZengNature PortfolioScientific Reports2045-23222023-01-0113112010.1038/s41598-023-27520-8Domain motions, dimerization, and membrane interactions of the murine guanylate binding protein 2Jennifer Loschwitz0Nora Steffens1Xue Wang2Moritz Schäffler3Klaus Pfeffer4Daniel Degrandi5Birgit Strodel6Institute of Theoretical and Computational Chemistry, Heinrich Heine University DüsseldorfInstitute of Medical Microbiology and Hospital Hygiene, Heinrich Heine University DüsseldorfInstitute of Theoretical and Computational Chemistry, Heinrich Heine University DüsseldorfInstitute of Biological Information Processing: Structural Biochemistry (IBI-7), Forschungszentrum JülichInstitute of Medical Microbiology and Hospital Hygiene, Heinrich Heine University DüsseldorfInstitute of Medical Microbiology and Hospital Hygiene, Heinrich Heine University DüsseldorfInstitute of Theoretical and Computational Chemistry, Heinrich Heine University DüsseldorfAbstract Guanylate-binding proteins (GBPs) are a group of GTPases that are induced by interferon- $$\gamma$$ γ and are crucial components of cell-autonomous immunity against intracellular pathogens. Here, we examine murine GBP2 (mGBP2), which we have previously shown to be an essential effector protein for the control of Toxoplasma gondii replication, with its recruitment through the membrane of the parasitophorous vacuole and its involvement in the destruction of this membrane likely playing a role. The overall aim of our work is to provide a molecular-level understanding of the mutual influences of mGBP2 and the parasitophorous vacuole membrane. To this end, we performed lipid-binding assays which revealed that mGBP2 has a particular affinity for cardiolipin. This observation was confirmed by fluorescence microscopy using giant unilamellar vesicles of different lipid compositions. To obtain an understanding of the protein dynamics and how this is affected by GTP binding, mGBP2 dimerization, and membrane binding, assuming that each of these steps are relevant for the function of the protein, we carried out standard as well as replica exchange molecular dynamics simulations with an accumulated simulation time of more than 30 μs. The main findings from these simulations are that mGBP2 features a large-scale hinge motion in its M/E domain, which is present in each of the studied protein states. When bound to a cardiolipin-containing membrane, this hinge motion is particularly pronounced, leading to an up and down motion of the M/E domain on the membrane, which did not occur on a membrane without cardiolipin. Our prognosis is that this up and down motion has the potential to destroy the membrane following the formation of supramolecular mGBP2 complexes on the membrane surface.https://doi.org/10.1038/s41598-023-27520-8 |
| spellingShingle | Jennifer Loschwitz Nora Steffens Xue Wang Moritz Schäffler Klaus Pfeffer Daniel Degrandi Birgit Strodel Domain motions, dimerization, and membrane interactions of the murine guanylate binding protein 2 Scientific Reports |
| title | Domain motions, dimerization, and membrane interactions of the murine guanylate binding protein 2 |
| title_full | Domain motions, dimerization, and membrane interactions of the murine guanylate binding protein 2 |
| title_fullStr | Domain motions, dimerization, and membrane interactions of the murine guanylate binding protein 2 |
| title_full_unstemmed | Domain motions, dimerization, and membrane interactions of the murine guanylate binding protein 2 |
| title_short | Domain motions, dimerization, and membrane interactions of the murine guanylate binding protein 2 |
| title_sort | domain motions dimerization and membrane interactions of the murine guanylate binding protein 2 |
| url | https://doi.org/10.1038/s41598-023-27520-8 |
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