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A pH induced two-dimensional crystal of membrane-bound Na+,K+-ATPase of dog kidney

Authors
Journal
FEBS Letters
0014-5793
Publisher
Wiley Blackwell (John Wiley & Sons)
Publication Date
Volume
320
Issue
1
Identifiers
DOI: 10.1016/0014-5793(93)81648-j
Keywords
  • Sodium Potassium Adenosine Triphosphatase (Na+
  • K+-Atpase)
  • Two-Dimensional Crystallization
  • Ice-Embedding Technique
  • Electron Cryo-Microscopy
  • Computer Assisted Image Processing

Abstract

Abstract Two-dimensional crystals of membrane-bound Na +,K +-ATPase were formed in acidic media and their qualities were investigated by electron cryo-microscopy as well as by conventional electron microscopy. At pH 4.8 in sodium citrate buffer, the best crystallization condition, more than 80% of membranes formed crystals. The high ratio allowed high-resolution images to be taken by electron cryo-microscopy. Image processing revealed that they had unique lattice constants ( a = 108.7 Å, b = 66.2 Å, γ = 104.2°) and had few defects in the crystalline arrays. The reconstituted Fourier map of the ice-embedded crystal showed that there are two high contrast parts in one unit cell.

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