TY - JOUR
T1 - Resolution of the V1 ATPase from Manduca sexta into subcomplexes and visualization of an ATPase-active A3B3EG complex by electron microscopy
AU - Rizzo, Vincenzo F.
AU - Coskun, Ünal
AU - Radermacher, Michael
AU - Ruiz, Teresa
AU - Armbrüster, Andrea
AU - Grüber, Gerhard
PY - 2003/1/3
Y1 - 2003/1/3
N2 - The effect of the ATPase activity of Manduca sexta V1 ATPase by the amphipathic detergent lauryldimethylamine oxide (LDAO) and the relationship of these activities to the subunit composition of V1 were studied. The V1 was highly activated in the presence of 0.04-0.06% LDAO combined with release of the subunits H, C, and F from the enzyme. Increase of LDAO concentration to 0.1-0.2% caused the characterized subcomplexes A3B3HEGF and A3B3EG with a remaining ATPase activity of 52 and 65%, respectively. The hydrolytic-active A3B3EG subcomplex has been visualized by electron microscopy showing six major masses of density in a pseudo-hexagonal arrangement surrounding a seventh mass. The compositions of the various subcomplexes and fragments of V1 provide an organization of the subunits in the enzyme in the framework of the known three-dimensional reconstruction of the V1 ATPase from M. sexta (Radermacher, M., Ruiz, T., Wieczorek, H., and Grüber, G. (2001) J. Struct. Biol. 135, 26-37).
AB - The effect of the ATPase activity of Manduca sexta V1 ATPase by the amphipathic detergent lauryldimethylamine oxide (LDAO) and the relationship of these activities to the subunit composition of V1 were studied. The V1 was highly activated in the presence of 0.04-0.06% LDAO combined with release of the subunits H, C, and F from the enzyme. Increase of LDAO concentration to 0.1-0.2% caused the characterized subcomplexes A3B3HEGF and A3B3EG with a remaining ATPase activity of 52 and 65%, respectively. The hydrolytic-active A3B3EG subcomplex has been visualized by electron microscopy showing six major masses of density in a pseudo-hexagonal arrangement surrounding a seventh mass. The compositions of the various subcomplexes and fragments of V1 provide an organization of the subunits in the enzyme in the framework of the known three-dimensional reconstruction of the V1 ATPase from M. sexta (Radermacher, M., Ruiz, T., Wieczorek, H., and Grüber, G. (2001) J. Struct. Biol. 135, 26-37).
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U2 - 10.1074/jbc.M208623200
DO - 10.1074/jbc.M208623200
M3 - Article
C2 - 12414800
AN - SCOPUS:0037414640
SN - 0021-9258
VL - 278
SP - 270
EP - 275
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 1
ER -