Three-dimensional organization of the archaeal A1-ATPase from methanosarcina mazei Gö1
Publikation: Beitrag in Fachzeitschrift › Forschungsartikel › Beigetragen › Begutachtung
Beitragende
Abstract
A modified isolation procedure provides a homogeneous A1-ATPase from the archaeon Methanosarcina mazei Gö1, containing the five subunits in stoichiometric amounts of A3:B3:C:D:F. A1 obtained in this way was characterized by three-dimensional electron microscopy of single particles, resulting in the first three-dimensional reconstruction of an A1-ATPase at a resolution of 3.2 nm. The A1 consists of a headpiece of 10.2 nm in diameter and 10.8 nm in height, formed by the six elongated subunits A3 and B3. At the bottom of the A 3B3 complex, a stalk of 3.0 nm in length can be seen. The A3B3 domain surrounds a large cavity that extends throughout the length of the A3B3 barrel. A part of the stalk penetrates inside this cavity and is displaced toward an A-B-A triplet. To investigate further the topology of the stalk subunits C-F in A1, cross-linking has been carried out by using dithiobis[sulfosuccinimidylpropionate] (DSP) and 1-ethyl-3-(dimethylaminopropyl)-carbodiimide (EDC). In experiments where DSP was added the cross-linked products B-F, Ax-D, A-B-D, and A x-Bx-D were formed. Subunits B-F, A-D, A-B-D, and A-B-C-D could be cross-linked by EDC. The subunit-subunit interaction in the presence of DSP was also studied as a function of nucleotide binding, demonstrating movements of subunits C, D, and F during ATP cleavage. Finally, the three-dimensional organization of this A1 complex is discussed in terms of the relationship to the F1- and V1-ATPases at a resolution of 3.2 nm.
Details
Originalsprache | Englisch |
---|---|
Seiten (von - bis) | 22759-22764 |
Seitenumfang | 6 |
Fachzeitschrift | Journal of Biological Chemistry |
Jahrgang | 279 |
Ausgabenummer | 21 |
Publikationsstatus | Veröffentlicht - 21 Mai 2004 |
Peer-Review-Status | Ja |
Extern publiziert | Ja |
Externe IDs
WOS | 000221417100127 |
---|---|
Scopus | 2542466633 |
PubMed | 14988401 |