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Updated: Sep 30, 2026

Optimized Negative Staining: a High-throughput Protocol for Examining Small and Asymmetric Protein Structure by Electron Microscopy
Published on: August 15, 2014
Projection structure of P-glycoprotein by electron microscopy. Evidence for a closed conformation of the nucleotide
Jyh-Yeuan Lee1, Ina L Urbatsch, Alan E Senior
1University of California, Riverside, Department of Biochemistry, Riverside, California 92521, USA.
Abstract:
The structure of P-glycoprotein (Pgp) from mouse has been studied by electron microscopy and image analysis. Two-dimensional crystals of Pgp in a lipid bilayer were generated by reconstituting pure, detergent-solubilized protein containing a C-terminal six-histidine tag using the lipid monolayer technique. The crystals belong to plane group P1 with a = b = 104 +/- 2 A and gamma = 90 +/- 4 degrees. The projection structure of Pgp calculated at a resolution of 22 A shows two closely interacting protein domains that can be interpreted as the N- and C-terminal halves of the protein. The projection structure of Pgp is consistent with the recently published x-ray structure of MsbA, a lipid A flippase from Escherichia coli with high sequence homology to Pgp but only when the two MsbA subunits are rotated to bring their nucleotide binding domains together.
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