HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 21, 22759-22764, May 21, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/21/22759    most recent M313741200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Coskun, U. Articles by Grüber, G. Search for Related Content PubMed PubMed Citation Articles by Coskun, U. Articles by Grüber, G. Social Bookmarking                      What's this?

Three-dimensional Organization of the Archaeal A₁-ATPase from Methanosarcina mazei Gö1^*

Ünal Coskun, Michael Radermacher, Volker Müller¶||, Teresa Ruiz¶, and Gerhard Grüber**

From the Universität des Saarlandes, Fachrichtung 2.5-Biophysik, D-66421 Homburg, Germany, the Department of Molecular Physiology and Biophysics, College of Medicine, University of Vermont, Burlington, Vermont 05405, and ^¶Ludwig Maximiliano Universität München, Department Biology I, Section Microbiology, Maria-Ward-Strasse 1a, D-80638 München, Germany

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 A₃B₃ 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, A_x-D, A-B-D, and A_x-B_x-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.

Received for publication, December 16, 2003 , and in revised form, February 20, 2004.

^* This work was supported by Grant GR 1475/9-1, 9-2 from the Deutsche Forschungsgemeinschaft. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| Present address: Johann Wolfgang Goethe-Universität Frankfurt, Institut für Mikrobiologie, D-60439 Frankfurt, Germany.

^** To whom correspondence should be addressed: Universität des Saarlandes, Fachrichtung 2.5-Biophysik, Universitätsbau 76, D-66421 Homburg, Germany. Tel.: 49-6841-162-6085; Fax: 49-6841-162-6086; E-mail: ggrueber{at}uniklinik-saarland.de.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				J. Vonck, K. Y. Pisa, N. Morgner, B. Brutschy, and V. Muller Three-dimensional Structure of A1A0 ATP Synthase from the Hyperthermophilic Archaeon Pyrococcus furiosus by Electron Microscopy J. Biol. Chem., April 10, 2009; 284(15): 10110 - 10119. [Abstract] [Full Text] [PDF]

				U. Coskun, Y. L. Chaban, A. Lingl, V. Muller, W. Keegstra, E. J. Boekema, and G. Gruber Structure and Subunit Arrangement of the A-type ATP Synthase Complex from the Archaeon Methanococcus jannaschii Visualized by Electron Microscopy J. Biol. Chem., September 10, 2004; 279(37): 38644 - 38648. [Abstract] [Full Text] [PDF]