Abstract
In spite of considerable progress in the methodology for reconstitution of membrane proteins into the liposomes, a successful reconstitution still appears to be more an art than a science. Reconstitution of membrane proteins into bilayers is required for establishing several aspects of the functions of membrane proteins and lipids and for elaborating models of naturally occurring membranes.
Cyclooxygenase (COX)-1 and -2 (also prostaglandin endoperoxide H2 synthase, PGHS-1 and -2) belong to the class of monotopic membrane proteins. Membrane-binding domains of both COX-1 and -2 contain four short, consecutive, amphipathic α-helices (A, B, C, and D). Crystal structures of the COXs indicate that basic, hydrophobic, and aromatic residues in the membrane-binding domain are oriented away from the protein core and form a surface on the enzyme, which has been proposed to interact with the lipid bilayer (1).
In this chapter, we describe a fast and efficient method for direct incorporation of COX-1 and -2 isozymes - as models for monotopic integral membrane proteins - into preformed liposomes containing fatty acids without loss of activity.
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References
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Acknowledgments
The authors would like to thank Dr. Alicia Pastor and Mr. Robert Pcionek from Michigan State University Center for Advanced Microscopy for their help with the electron microscopy work.
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MirAfzali, Z., DeWitt, D.L. (2010). Liposomal Reconstitution of Monotopic Integral Membrane Proteins. In: Weissig, V. (eds) Liposomes. Methods in Molecular Biology™, vol 606. Humana Press. https://doi.org/10.1007/978-1-60761-447-0_7
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DOI: https://doi.org/10.1007/978-1-60761-447-0_7
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