Three-dimensional structure of the KdpFABC complex of Escherichia coli by electron tomography of two-dimensional crystals
Autor(en): | Hu, Guo-Bin Rice, William J. Droese, Stefan Altendorf, Karlheinz Stokes, David L. |
Stichwörter: | AMINO-ACID SUBSTITUTIONS; Biochemistry & Molecular Biology; Biophysics; CALCIUM-PUMP; Cell Biology; electron tomography; homology model; ion transport; K+-TRANSPORT; Kdp; KDP-ATPASE; P-type ATPase; P-TYPE ATPASES; real-space average; RECONSTRUCTION; SARCOPLASMIC-RETICULUM; SUBUNIT; TRANSPORT ATPASE; VISUALIZATION | Erscheinungsdatum: | 2008 | Herausgeber: | ACADEMIC PRESS INC ELSEVIER SCIENCE | Journal: | JOURNAL OF STRUCTURAL BIOLOGY | Volumen: | 161 | Ausgabe: | 3 | Startseite: | 411 | Seitenende: | 418 | Zusammenfassung: | The KdpFABC complex (Kdp) functions as a K+ pump in Escherichia coli and is a member of the family of P-type ATPases. Unlike other family members, Kdp has a unique oligomeric composition and is notable for segregating K+ transport and ATP hydrolysis onto separate subunits (KdpA and KdpB, respectively). We have produced two-dimensional crystals of the KdpFABC complex within reconstituted lipid bilayers and determined its three-dimensional structure from negatively stained samples using a combination of electron tomography and real-space averaging. The resulting map is at a resolution of 2.4 nm and reveals a dimer of Kdp molecules as the asymmetric unit; however, only the cytoplasmic domains are visible due to the lack of stain penetration within the lipid bilayer. The sizes of these cytoplasmic domains are consistent with Kdp and, using a pseudo-atomic model, we have described the subunit interactions that stabilize the Kdp dimer within the larger crystallographic array. These results illustrate the utility of electron tomography in structure determination of ordered assemblies, especially when disorder is severe enough to hamper conventional crystallographic analysis. (C) 2007 Elsevier Inc. All rights reserved. |
Beschreibung: | 4th International Conference on Electron Tomography, San Diego, CA, NOV 05-08, 2006 |
ISSN: | 10478477 | DOI: | 10.1016/j.jsb.2007.09.006 |
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