TY - JOUR
T1 - Mitochondrial ATP synthasome. Three-dimensional structure by electron microscopy of the ATP synthase in complex formation with carriers for P i and ADP/ATP
AU - Chen, Chen
AU - Ko, Young
AU - Delannoy, Michael
AU - Ludtke, Steven J.
AU - Chiu, Wah
AU - Pedersen, Peter L.
PY - 2004/7/23
Y1 - 2004/7/23
N2 - The terminal steps involved in making ATP in mitochondria require an ATP synthase (F0F1) comprised of two motors, a phosphate carrier (PIC), and an adenine nucleotide carrier (ANC). Under mild conditions, these entities sub-fractionate as an ATP synthase/PIC/ANC complex or "ATP synthasome" (Ko, Y.H., Delannoy, M., Hullihen, J., Chiu, W., and Pedersen, P.L. (2003) J. Biol. Chem. 278, 12305-12309). As a first step toward obtaining three-dimensional information about this large complex or "metabolon" and the locations of PIC and ANC therein, we dispersed ATP synthasomes into single complexes and visualized negatively stained images by electron microscopy (EM) that showed clearly the classical headpiece, central stalk, and basepiece. Parallel immuno-EM studies revealed the presence of PIC and ANC located non-centrally in the basepiece, and other studies implicated an ATP synthase/PIC/ANC stoichiometry near 1:1:1. Single ATP synthasome images (7506) were boxed, and, using EMAN software, a three-dimensional model was obtained at a resolution of 23 Å. Significantly, the basepiece is oblong and contains two domains, the larger of which connects to the central stalk, whereas the smaller appears as an extension. Docking studies with known structures together with the immuno-EM studies suggest that PIC or ANC may be located in the smaller domain, whereas the other transporter resides nearby in the larger domain. Collectively, these finding support a mechanism in which the entry of the substrates ADP and Pi into mitochondria, the synthesis of ATP on F1, and the release and exit of ATP are very localized and highly coordinated events.
AB - The terminal steps involved in making ATP in mitochondria require an ATP synthase (F0F1) comprised of two motors, a phosphate carrier (PIC), and an adenine nucleotide carrier (ANC). Under mild conditions, these entities sub-fractionate as an ATP synthase/PIC/ANC complex or "ATP synthasome" (Ko, Y.H., Delannoy, M., Hullihen, J., Chiu, W., and Pedersen, P.L. (2003) J. Biol. Chem. 278, 12305-12309). As a first step toward obtaining three-dimensional information about this large complex or "metabolon" and the locations of PIC and ANC therein, we dispersed ATP synthasomes into single complexes and visualized negatively stained images by electron microscopy (EM) that showed clearly the classical headpiece, central stalk, and basepiece. Parallel immuno-EM studies revealed the presence of PIC and ANC located non-centrally in the basepiece, and other studies implicated an ATP synthase/PIC/ANC stoichiometry near 1:1:1. Single ATP synthasome images (7506) were boxed, and, using EMAN software, a three-dimensional model was obtained at a resolution of 23 Å. Significantly, the basepiece is oblong and contains two domains, the larger of which connects to the central stalk, whereas the smaller appears as an extension. Docking studies with known structures together with the immuno-EM studies suggest that PIC or ANC may be located in the smaller domain, whereas the other transporter resides nearby in the larger domain. Collectively, these finding support a mechanism in which the entry of the substrates ADP and Pi into mitochondria, the synthesis of ATP on F1, and the release and exit of ATP are very localized and highly coordinated events.
UR - http://www.scopus.com/inward/record.url?scp=3843147327&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=3843147327&partnerID=8YFLogxK
U2 - 10.1074/jbc.M401353200
DO - 10.1074/jbc.M401353200
M3 - Article
C2 - 15166242
AN - SCOPUS:3843147327
SN - 0021-9258
VL - 279
SP - 31761
EP - 31768
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 30
ER -