Solution structure of the KdpFABC P-type ATPase from Escherichia coli by electron microscopic single particle analysis

Autor(en): Heitkamp, Thomas
Boettcher, Bettina
Greie, Joerg-Christian
Stichwörter: 2-DIMENSIONAL CRYSTALS; AMINO-ACID SUBSTITUTIONS; Biochemistry & Molecular Biology; Biophysics; CALCIUM-PUMP; Cell Biology; COMPLEX; CRYSTAL-STRUCTURE; Electron microscopy; ION-TRANSPORT; K+-BINDING; KDP-ATPASE; KdpFABC; P-type ATPase; Potassium transport; Single particle analysis; TRANSPORT ATPASE; UNIQUE FEATURES
Erscheinungsdatum: 2009
Herausgeber: ACADEMIC PRESS INC ELSEVIER SCIENCE
Journal: JOURNAL OF STRUCTURAL BIOLOGY
Volumen: 166
Ausgabe: 3
Startseite: 295
Seitenende: 302
Zusammenfassung: 
The K+-translocating KdpFABC complex from Escherichia coli functions as a high affinity potassium uptake system and belongs to the superfamily of P-type ATPases, although it exhibits some unique features. It comprises four subunits, and the sites of ATP hydrolysis and substrate transport are located on two different polypeptides. No structural data are so far available for elucidating the correspondingly unique mechanism of coupling ion transport and catalysis in this P-type ATPase. By use of electron microscopy and single particle analysis of negatively stained, solubilized KdpFABC complexes, we solved the structure of the complex at a resolution of 19 A, which allowed us to model the arrangement of subunits within the holoenzyme and, thus, to identify the interfaces between subunits. The model showed that the K+-translocating KdpA subunit is in close contact with the transmembrane region of the ATP-hydrolyzing subunit KdpB. The cytosolic C-terminal domain of the KdpC subunit, which is assumed to play a role in cooperative ATP binding together with KdpB, is located in close vicinity to the nucleotide binding domain of KdpB. Overall, the arrangement of subunits agrees with biochemical data and the predictions on subunit interactions. (C) 2009 Elsevier Inc. All rights reserved.
ISSN: 10478477
DOI: 10.1016/j.jsb.2009.02.016

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