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Science 20 September 2002:
Vol. 297. no. 5589, pp. 2018 - 2026
DOI: 10.1126/science.1074424
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Research Articles

Nucleotide Control of Interdomain Interactions in the Conformational Reaction Cycle of SecA

John F. Hunt,12*dagger Sevil Weinkauf,23* Lisa Henry,2 John J. Fak,1* Paul McNicholas,4* Donald B. Oliver,4 Johann Deisenhofer2

The SecA adenosine triphosphatase (ATPase) mediates extrusion of the amino termini of secreted proteins from the eubacterial cytosol based on cycles of reversible binding to the SecYEG translocon. We have determined the crystal structure of SecA with and without magnesium-adenosine diphosphate bound to the high-affinity ATPase site at 3.0 and 2.7 angstrom resolution, respectively. Candidate sites for preprotein binding are located on a surface containing the SecA epitopes exposed to the periplasm upon binding to SecYEG and are thus positioned to deliver preprotein to SecYEG. Comparisons with structurally related ATPases, including superfamily I and II ATP-dependent helicases, suggest that the interaction geometry of the tandem motor domains in SecA is modulated by nucleotide binding, which is shown by fluorescence anisotropy experiments to reverse an endothermic domain-dissociation reaction hypothesized to gate binding to SecYEG.

1 Department of Biological Sciences, 702A Fairchild Center, MC2434, Columbia University, New York, NY 10027, USA.
2 Howard Hughes Medical Institute and Department of Biochemistry, The University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, TX 75390-9050, USA.
3 Department of Chemistry, Technical University Munich, Lichtenbergstrasse 4, D-85748 Garching, Germany.
4 Department of Molecular Biology and Biochemistry, Lawn Avenue, Wesleyan University, Middletown, CT 06459, USA.
*   Present addresses: Columbia University, New York, NY 10027, USA. (J.F.H.); Technical University Munich, Lichtenbergstrasse 4, D-85748 Garching, Germany (S.W.); Laboratory of Developmental Neurogenetics, Rockefeller University, New York, NY 10021, USA (J.J.F.); Schering-Plough Research Institute, Kenilworth, NJ 07033, USA (P.M.).

dagger    To whom correspondence should be addressed. E-mail: hunt{at}sid.bio.columbia.edu

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P. Natale, J. Swaving, C. van der Does, J. de Keyzer, and A. J. M. Driessen (2004)
J. Biol. Chem. 279, 13769-13777
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