Electrostatic Forces Mediate the Specificity of RHO GTPase-GDI Interactions
Three decades of research have documented the spatiotemporal dynamics of RHO family GTPase membrane extraction regulated by guanine nucleotide dissociation inhibitors (GDIs), but the interplay of the kinetic mechanism and structural specificity of these interactions is as yet unresolved. To address...
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MDPI AG
2021-11-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/22/12493 |
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| author | Niloufar Mosaddeghzadeh Neda S. Kazemein Jasemi Jisca Majolée Si-Cai Zhang Peter L. Hordijk Radovan Dvorsky Mohammad Reza Ahmadian |
| author_facet | Niloufar Mosaddeghzadeh Neda S. Kazemein Jasemi Jisca Majolée Si-Cai Zhang Peter L. Hordijk Radovan Dvorsky Mohammad Reza Ahmadian |
| author_sort | Niloufar Mosaddeghzadeh |
| collection | DOAJ |
| description | Three decades of research have documented the spatiotemporal dynamics of RHO family GTPase membrane extraction regulated by guanine nucleotide dissociation inhibitors (GDIs), but the interplay of the kinetic mechanism and structural specificity of these interactions is as yet unresolved. To address this, we reconstituted the GDI-controlled spatial segregation of geranylgeranylated RHO protein RAC1 in vitro. Various biochemical and biophysical measurements provided unprecedented mechanistic details for GDI function with respect to RHO protein dynamics. We determined that membrane extraction of RHO GTPases by GDI occurs via a 3-step mechanism: (1) GDI non-specifically associates with the switch regions of the RHO GTPases; (2) an electrostatic switch determines the interaction specificity between the C-terminal polybasic region of RHO GTPases and two distinct negatively-charged clusters of GDI1; (3) a non-specific displacement of geranylgeranyl moiety from the membrane sequesters it into a hydrophobic cleft, effectively shielding it from the aqueous milieu. This study substantially extends the model for the mechanism of GDI-regulated RHO GTPase extraction from the membrane, and could have implications for clinical studies and drug development. |
| first_indexed | 2024-03-10T05:25:13Z |
| format | Article |
| id | doaj.art-016abdbc02aa459eb551752890f5a0da |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T05:25:13Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-016abdbc02aa459eb551752890f5a0da2023-11-22T23:43:19ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-11-0122221249310.3390/ijms222212493Electrostatic Forces Mediate the Specificity of RHO GTPase-GDI InteractionsNiloufar Mosaddeghzadeh0Neda S. Kazemein Jasemi1Jisca Majolée2Si-Cai Zhang3Peter L. Hordijk4Radovan Dvorsky5Mohammad Reza Ahmadian6Institute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, 40225 Düsseldorf, GermanyInstitute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, 40225 Düsseldorf, GermanyDepartment of Physiology, Amsterdam UMC, Location VUmc, De Boelelaan, 1108 Amsterdam, The NetherlandsInstitute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, 40225 Düsseldorf, GermanyDepartment of Physiology, Amsterdam UMC, Location VUmc, De Boelelaan, 1108 Amsterdam, The NetherlandsInstitute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, 40225 Düsseldorf, GermanyInstitute of Biochemistry and Molecular Biology II, Medical Faculty and University Hospital Düsseldorf, Heinrich Heine University Düsseldorf, 40225 Düsseldorf, GermanyThree decades of research have documented the spatiotemporal dynamics of RHO family GTPase membrane extraction regulated by guanine nucleotide dissociation inhibitors (GDIs), but the interplay of the kinetic mechanism and structural specificity of these interactions is as yet unresolved. To address this, we reconstituted the GDI-controlled spatial segregation of geranylgeranylated RHO protein RAC1 in vitro. Various biochemical and biophysical measurements provided unprecedented mechanistic details for GDI function with respect to RHO protein dynamics. We determined that membrane extraction of RHO GTPases by GDI occurs via a 3-step mechanism: (1) GDI non-specifically associates with the switch regions of the RHO GTPases; (2) an electrostatic switch determines the interaction specificity between the C-terminal polybasic region of RHO GTPases and two distinct negatively-charged clusters of GDI1; (3) a non-specific displacement of geranylgeranyl moiety from the membrane sequesters it into a hydrophobic cleft, effectively shielding it from the aqueous milieu. This study substantially extends the model for the mechanism of GDI-regulated RHO GTPase extraction from the membrane, and could have implications for clinical studies and drug development.https://www.mdpi.com/1422-0067/22/22/12493CDC42electrostatic steeringG domainhypervariable regiongeranylgeranylguanine nucleotide dissociation inhibitors |
| spellingShingle | Niloufar Mosaddeghzadeh Neda S. Kazemein Jasemi Jisca Majolée Si-Cai Zhang Peter L. Hordijk Radovan Dvorsky Mohammad Reza Ahmadian Electrostatic Forces Mediate the Specificity of RHO GTPase-GDI Interactions International Journal of Molecular Sciences CDC42 electrostatic steering G domain hypervariable region geranylgeranyl guanine nucleotide dissociation inhibitors |
| title | Electrostatic Forces Mediate the Specificity of RHO GTPase-GDI Interactions |
| title_full | Electrostatic Forces Mediate the Specificity of RHO GTPase-GDI Interactions |
| title_fullStr | Electrostatic Forces Mediate the Specificity of RHO GTPase-GDI Interactions |
| title_full_unstemmed | Electrostatic Forces Mediate the Specificity of RHO GTPase-GDI Interactions |
| title_short | Electrostatic Forces Mediate the Specificity of RHO GTPase-GDI Interactions |
| title_sort | electrostatic forces mediate the specificity of rho gtpase gdi interactions |
| topic | CDC42 electrostatic steering G domain hypervariable region geranylgeranyl guanine nucleotide dissociation inhibitors |
| url | https://www.mdpi.com/1422-0067/22/22/12493 |
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