Abstract：
: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.
journal_name
J Struct Bioljournal_title
Journal of structural biologyauthors
Hu GB,Rice WJ,Dröse S,Altendorf K,Stokes DLdoi
10.1016/j.jsb.2007.09.006subject
Has Abstractpub_date
2008-03-01 00:00:00pages
411-8issue
3eissn
1047-8477issn
1095-8657pii
S1047-8477(07)00223-7journal_volume
161pub_type
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journal_title：Journal of structural biology
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journal_title：Journal of structural biology
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journal_title：Journal of structural biology
pub_type： 杂志文章,评审
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