title: Integrating three views of Arf1 activation dynamics.
abstract: The proteins Arno and Gea2 of the Sec7 family can promote GDP-GTP exchange on Arf1, a small GTP-binding protein, which coordinates coated vesicle formation for protein transport within the cell. Crystal structures of the essential Sec7 domain (Sec7d) of Gea2 in the free and Arf1-bound forms suggest that conformational dynamics of the Sec7d as well as those of the G-protein play a role in nucleotide exchange. Starting from a set of complementary crystal structures, we compared the collective movements of unbound Gea2 and Arno Sec7 domains, Arf1-GDP, and the Arf1-Gea2(Sec7d) nucleotide-free complex using normal modes analyses. In all unbound Sec7d analyses, significant low-energy movements were found to lead to closure of the hydrophobic groove towards the form seen in the Arf1-Gea2(Sec7d) complex, suggesting that groove closure is a general feature of the Sec7 family. Low-energy movements in Arf1-GDP implicate critical switch 1 and 2 residues which are coupled to modifications in the myristoylated N-terminal-helix binding site at the other end of the "interswitch" beta hairpin. It is suggested that Sec7d groove closure upon docking of the two molecules may permit extraction of switch 1 from Arf1-GDP and prepare the complex for movement of the interswitch, which is central to the membrane-linked exchange activity. Large-scale collective movements in the Arf1-Sec7d complex appear to participate in the insertion of the Sec7d Glu finger into the GDP binding site to promote actual nucleotide release.
authors.affiliation: Modélisation et Ingénierie des Protéines, CNRS UMR 8619, Bât. 430, Université de Paris-Sud 91405 Orsay, France.
authors.forename: Charles H
authors.initials: CH
authors.lastname: Robert
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Cherfils
authors.forename: Liliane
authors.initials: L
authors.lastname: Mouawad
authors.forename: David
authors.initials: D
authors.lastname: Perahia
mesh_terms: D020823:ADP-Ribosylation Factor 1 / Q000737:chemistry / Q000378:metabolism*; D004789:Enzyme Activation; D020690:GTPase-Activating Proteins / Q000378:metabolism; D020662:Guanine Nucleotide Exchange Factors / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: C121632:GEA2 protein, S cerevisiae; D020690:GTPase-Activating Proteins; D020662:Guanine Nucleotide Exchange Factors; D029701:Saccharomyces cerevisiae Proteins; C102474:cytohesin-2; D020823:ADP-Ribosylation Factor 1