title: Dual interaction of ADP ribosylation factor 1 with Sec7 domain and with lipid membranes during catalysis of guanine nucleotide exchange.
abstract: Sec7 domains catalyze the replacement of GDP by GTP on the G protein ADP-ribosylation factor 1 (myrARF1) by interacting with its switch I and II regions and by destabilizing, through a glutamic finger, the beta-phosphate of the bound GDP. The myristoylated N-terminal helix that allows myrARF1 to interact with membrane lipids in a GTP-dependent manner is located some distance from the Sec7 domain-binding region. However, these two regions are connected. Measuring the binding to liposomes of functional or abortive complexes between myrARF1 and the Sec7 domain of ARNO demonstrates that myrARF1, in complex with the Sec7 domain, adopts a high affinity state for membrane lipids, similar to that of the free GTP-bound form. This tight membrane attachment does not depend on the release of GDP induced by the Sec7 domain but is partially inhibited by the uncompetitive inhibitor brefeldin A. These results suggest that the conformational switch of the N-terminal helix of myrARF1 to the membrane-bound form is an early event in the nucleotide exchange pathway and is a prerequisite for a structural rearrangement at the myrARF1-GDP/Sec7 domain interface that allows the glutamic finger to expel GDP from myrARF1.
authors.affiliation: CNRS, Institut de Pharmacologie Moléculaire et Cellulaire, 660 route des lucioles, 06560 Valbonne, France.
authors.forename: S
authors.initials: S
authors.lastname: Béraud-Dufour
authors.forename: S
authors.initials: S
authors.lastname: Paris
authors.forename: M
authors.initials: M
authors.lastname: Chabre
authors.forename: B
authors.initials: B
authors.lastname: Antonny
mesh_terms: D020823:ADP-Ribosylation Factor 1 / Q000737:chemistry / Q000378:metabolism*; D002384:Catalysis; D005656:Fungal Proteins / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D008081:Liposomes; D008563:Membrane Lipids / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D005656:Fungal Proteins; D020662:Guanine Nucleotide Exchange Factors; D008081:Liposomes; D008563:Membrane Lipids; C066755:Sec7 guanine nucleotide exchange factors; D020823:ADP-Ribosylation Factor 1