Crystal structure of ARF1*Sec7 complexed with Brefeldin A and its implications for the guanine nucleotide exchange mechanism.
ARF GTPases are activated by guanine nucleotide exchange factors (GEFs) of the Sec7 family that promote the exchange of GDP for GTP. Brefeldin A (BFA) is a fungal metabolite that binds to the ARF1*GDP*Sec7 complex and blocks GEF activity at an early stage of the reaction, prior to guanine nucleotide release. The crystal structure of the ARF1*GDP*Sec7*BFA complex shows that BFA binds at the protein-protein interface to inhibit conformational changes in ARF1 required for Sec7 to dislodge the GDP molecule. Based on a comparative analysis of the inhibited complex, nucleotide-free ARF1*Sec7 and ARF1*GDP, we suggest that, in addition to forcing nucleotide release, the ARF1-Sec7 binding energy is used to open a cavity on ARF1 to facilitate the rearrangement of hydrophobic core residues between the GDP and GTP conformations. Thus, the Sec7 domain may act as a dual catalyst, facilitating both nucleotide release and conformational switching on ARF proteins.
Mossessova
Elena
E
Howard Hughes Medical Institute, Structural Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021, USA.
Corpina
Richard A
RA
Goldberg
Jonathan
J
D020823:ADP-Ribosylation Factor 1 / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D020126:Brefeldin A / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D004789:Enzyme Activation; D020662:Guanine Nucleotide Exchange Factors / Q000737:chemistry* / Q000378:metabolism; D006153:Guanosine Diphosphate / Q000737:chemistry / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances; D008958:Models, Molecular; D020836:Protein Structure, Quaternary; D011500:Protein Synthesis Inhibitors / Q000737:chemistry* / Q000378:metabolism
D020662:Guanine Nucleotide Exchange Factors; D046911:Macromolecular Substances; D011500:Protein Synthesis Inhibitors; C066755:Sec7 guanine nucleotide exchange factors; D006153:Guanosine Diphosphate; D020126:Brefeldin A; D006160:Guanosine Triphosphate; D020823:ADP-Ribosylation Factor 1