Projection structure of the atractyloside-inhibited mitochondrial ADP/ATP carrier of Saccharomyces cerevisiae.
ADP/ATP carriers in the inner mitochondrial membrane catalyze the exchange of cytosolic ADP for ATP synthesized in the mitochondrial matrix by ATP synthase and thereby replenish the eukaryotic cell with metabolic energy. The yeast ADP/ATP carrier (AAC3) was overexpressed, inhibited by atractyloside, purified, and reconstituted into two-dimensional crystals. Images of frozen hydrated crystals were recorded by electron microscopy, and a projection structure was calculated to 8-A resolution. The AAC3 molecule has pseudo 3-fold symmetry in agreement with the 3-fold sequence repeats that are typical of members of the mitochondrial carrier family. The density distribution is consistent with a bundle of six transmembrane alpha-helices with two or three short alpha-helical extensions closing the central pore on the matrix side. The AAC3 molecules in the crystal are arranged in symmetrical homo-dimers, but the translocation pore for adenine nucleotides lies in the center of the molecule and not along the dyad axis of the dimer.
Kunji
Edmund R S
ER
Medical Research Council, Dunn Human Nutrition Unit, Hills Road, Cambridge CB2 2XY, United Kingdom. ek@mrc-dunn.cam.ac.uk
Harding
Marilyn
M
D001278:Atractyloside / Q000494:pharmacology*; D003461:Crystallography; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D000226:Mitochondrial ADP, ATP Translocases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*
D004791:Enzyme Inhibitors; D029701:Saccharomyces cerevisiae Proteins; D001278:Atractyloside; D000226:Mitochondrial ADP, ATP Translocases