Kinetics of interaction between ADP-ribosylation factor-1 (Arf1) and the Sec7 domain of Arno guanine nucleotide exchange factor, modulation by allosteric factors, and the uncompetitive inhibitor brefeldin A
| Autor: | Martine Pugnière, Sana Bakari, Sebastien Estaran, Alain Chavanieu, Jad Rouhana, Stephane Delbecq, André Padilla, Joël Chopineau, Yvan Boublik |
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| Přispěvatelé: | Institut des Biomolécules Max Mousseron [Pôle Chimie Balard] (IBMM), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM), Centre de Biochimie Structurale [Montpellier] (CBS), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM), Vaccination Antiparasitaire : Laboratoire de Biologie Cellulaire et Moléculaire (LBCM), Université Montpellier 1 (UM1)-Université de Montpellier (UM), Centre de recherche en Biologie Cellulaire (CRBM), Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 1 (UM1), Institut Charles Gerhardt Montpellier - Institut de Chimie Moléculaire et des Matériaux de Montpellier (ICGM ICMMM), Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM)-Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut de Chimie du CNRS (INC), Institut de recherche en cancérologie de Montpellier (IRCM - U896 Inserm - UM1), Université Montpellier 1 (UM1)-CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM) |
| Rok vydání: | 2012 |
| Předmět: |
GTP'
Stereochemistry Allosteric regulation [SDV.BC]Life Sciences [q-bio]/Cellular Biology Guanosine triphosphate Biochemistry Binding Competitive Guanosine Diphosphate Catalysis Small gtpases 03 medical and health sciences chemistry.chemical_compound Escherichia coli Guanine Nucleotide Exchange Factors Humans Nucleotide Biotinylation Molecular Biology 030304 developmental biology chemistry.chemical_classification 0303 health sciences Brefeldin A organic chemicals 030302 biochemistry & molecular biology GTPase-Activating Proteins Cell Biology Surface Plasmon Resonance Kinetics chemistry Surface plasmon resonance (SPR) Guanosine diphosphate Guanine nucleotide exchange factor (GEF) ADP-Ribosylation Factor 1 Brefeldin A (BFA) Guanine nucleotide exchange factor Guanosine Triphosphate Uncompetitive inhibitor Allosteric Site Molecular Biophysics Plasmids Protein Binding |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2013, 288 (7), pp.4659-72. ⟨10.1074/jbc.M112.391748⟩ |
| ISSN: | 1083-351X 0021-9258 |
| Popis: | International audience; The GDP/GTP nucleotide exchange of Arf1 is catalyzed by nucleotide exchange factors (GEF), such as Arno, which act through their catalytic Sec7 domain. This exchange is a complex mechanism that undergoes conformational changes and intermediate complex species involving several allosteric partners such as nucleotides, Mg(2+), and Sec7 domains. Using a surface plasmon resonance approach, we characterized the kinetic binding parameters for various intermediate complexes. We first confirmed that both GDP and GTP counteract equivalently to the free-nucleotide binary Arf1-Arno complex stability and revealed that Mg(2+) potentiates by a factor of 2 the allosteric effect of GDP. Then we explored the uncompetitive inhibitory mechanism of brefeldin A (BFA) that conducts to an abortive pentameric Arf1-Mg(2+)-GDP-BFA-Sec7 complex. With BFA, the association rate of the abortive complex is drastically reduced by a factor of 42, and by contrast, the 15-fold decrease of the dissociation rate concurs to stabilize the pentameric complex. These specific kinetic signatures have allowed distinguishing the level and nature as well as the fate in real time of formed complexes according to experimental conditions. Thus, we showed that in the presence of GDP, the BFA-resistant Sec7 domain of Arno can also associate to form a pentameric complex, which suggests that the uncompetitive inhibition by BFA and the nucleotide allosteric effect combine to stabilize such abortive complex. |
| Databáze: | OpenAIRE |
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