A glutamic finger in the guanine nucleotide exchange factor ARNO displaces Mg2+ and the beta -phosphate to destabilize GDP on ARF1

Autor: Jacqueline Cherfils, Pierre Chardin, Marc Chabre, Sonia Paris, Bruno Antonny, Sophie Béraud-Dufour, Sylviane Robineau
Přispěvatelé: Institut de pharmacologie moléculaire et cellulaire (IPMC), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS), Unité des Rickettsies et pathogènes émergents (URPE), Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes (URMITE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR48, INSB-INSB-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR48, INSB-INSB-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'enzymologie et biochimie structurales (LEBS), Centre National de la Recherche Scientifique (CNRS), Université Nice Sophia Antipolis (1965 - 2019) (UNS), Institut des sciences biologiques (INSB-CNRS)-Institut des sciences biologiques (INSB-CNRS)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR48, Institut des sciences biologiques (INSB-CNRS)-Institut des sciences biologiques (INSB-CNRS)-Centre National de la Recherche Scientifique (CNRS)
Jazyk: angličtina
Rok vydání: 1998
Předmět:
Models
Molecular
ADP ribosylation factor
GTPase-activating protein
Protein Conformation
Stereochemistry
[SDV]Life Sciences [q-bio]
Glutamic Acid
Biology
Guanosine Diphosphate
General Biochemistry
Genetics and Molecular Biology
Phosphates
Fungal Proteins
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
GTP-binding protein regulators
Protein structure
GTP-Binding Proteins
Guanine Nucleotide Exchange Factors
Magnesium
[SDV.BBM]Life Sciences [q-bio]/Biochemistry
Molecular Biology
Binding site
Molecular Biology
ComputingMilieux_MISCELLANEOUS
030304 developmental biology
Aspartic Acid
0303 health sciences
Fungal protein
Binding Sites
General Immunology and Microbiology
ADP-Ribosylation Factors
General Neuroscience
GTPase-Activating Proteins
Biochemistry
chemistry
Guanosine diphosphate
Mutagenesis
Site-Directed
ADP-Ribosylation Factor 1
Guanine nucleotide exchange factor
030217 neurology & neurosurgery
Research Article
Zdroj: EMBO Journal
EMBO Journal, EMBO Press, 1998, 17 (13), pp.3651-3659. ⟨10.1093/emboj/17.13.3651⟩
EMBO Journal, 1998, 17 (13), pp.3651-3659. ⟨10.1093/emboj/17.13.3651⟩
ISSN: 0261-4189
1460-2075
DOI: 10.1093/emboj/17.13.3651⟩
Popis: The Sec7 domain of the guanine nucleotide exchange factor ARNO (ARNO-Sec7) is responsible for the exchange activity on the small GTP-binding protein ARF1. ARNO-Sec7 forms a stable complex with the nucleotide-free form of [Delta17]ARF1, a soluble truncated form of ARF1. The crystal structure of ARNO-Sec7 has been solved recently, and a site-directed mutagenesis approach identified a hydrophobic groove and an adjacent hydrophilic loop as the ARF1-binding site. We show that Glu156 in the hydrophilic loop of ARNO-Sec7 is involved in the destabilization of Mg2+ and GDP from ARF1. The conservative mutation E156D and the charge reversal mutation E156K reduce the exchange activity of ARNO-Sec7 by several orders of magnitude. Moreover, [E156K]ARNO-Sec7 forms a complex with the Mg2+-free form of [Delta17]ARF1-GDP without inducing the release of GDP. Other mutations in ARNO-Sec7 and in [Delta17]ARF1 suggest that prominent hydrophobic residues of the switch I region of ARF1 insert into the groove of the Sec7 domain, and that Lys73 of the switch II region of ARF1 forms an ion pair with Asp183 of ARNO-Sec7.
Databáze: OpenAIRE
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