Engineering Commons^™

Reconstitution And Characterization Of Membrane Proteins, Sarah Mia Pates

Masters Theses

“A membrane protein, cytochrome c reductase, was crystallized and examined using transmission electron microscopy. Two-dimensional crystals were prepared by two methods. In Method 1, the protein was solubilized in 50 mM Tris-Acetate (pH 6.0)/0.5 mM EDTA70.05% (w/v) Triton X-100 and reconstituted into a mixture of phosphatidylcholine and phosphatidylserine (4:1), dissolved in the same buffer. The Triton X-100 was removed from the mixture by dialysis in the presence of Bio-beads (SM-2) at approximately 25-30 °C. Method 2 was the same as Method 1, except that the vesicles were fractionated before reconstitution, and the pH and temperature for crystallization were 7.0 and …