Cryo-electron Microscopy of the Vacuolar ATPase Motor Reveals its Mechanical and Regulatory Complexity

Autor(en): Muench, Stephen P. 
Huss, Markus 
Song, Chun Feng
Phillips, Clair 
Wieczorek, Helmut 
Trinick, John 
Harrison, Michael A. 
Stichwörter: 3D reconstruction; Biochemistry & Molecular Biology; cryo-electron microscopy; CRYSTAL-STRUCTURE; ELECTRON-MICROSCOPY; ENTEROCOCCUS-HIRAE; H+-ATPASE; MEDIATED CROSS-LINKING; NA+-ATPASE; PERIPHERAL STALK; SACCHAROMYCES-CEREVISIAE; SUBUNIT-C VMA5P; vacuolar membrane; YEAST V-ATPASE
Erscheinungsdatum: 2009
Herausgeber: ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
Journal: JOURNAL OF MOLECULAR BIOLOGY
Volumen: 386
Ausgabe: 4
Startseite: 989
Seitenende: 999
Zusammenfassung: 
The vacuolar H+-ATPase (V-ATPase) is an ATP-driven rotary molecular motor that is a transmembrane proton pump in all eukaryotic cells. Although its activity is fundamental to many physiological processes, our understanding of the structure and mechanism of the V-ATPase is poor. Using cryo-electron microscopy of the tobacco hornworm (Manduca sexta) enzyme, we have calculated the first 3D reconstruction of the intact pump in its native state. The resolution of 16.5 angstrom is significantly higher than that of previous cryo-electron microscopy models of either V-ATPase or the related F1F0-ATPase. A network of four stalk structures connecting the V-1 catalytic domain and the V-0 membrane domain is now fully resolved, demonstrating substantially greater complexity than that found in the F-ATPase. Three peripheral stator stalks, connect those domains to a horizontal collar that partly encircles the region between V-1 and V-0. The fourth stalk is a central axle that connects to V-0 but makes minimal contact with V1. Several subunit crystal structures can be fit accurately into the reconstruction. The model thus provides new insights into the organisation of key components involved in mechanical coupling between the domains and regulation of activity. (C) 2009 Elsevier Ltd. All rights reserved.
ISSN: 00222836
DOI: 10.1016/j.jmb.2009.01.014

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