Liposomal reconstitution of monotopic integral membrane proteins.
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 H(2) 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 alpha-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.
MirAfzali
Zahra
Z
Encapsula NanoSciences LLC, Nashville, TN 37214, USA.
DeWitt
David L
DL
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002460:Cell Line; D051545:Cyclooxygenase 1 / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism; D015870:Gene Expression; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D008565:Membrane Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D018411:Spodoptera / Q000235:genetics
D005227:Fatty Acids; D008081:Liposomes; D008565:Membrane Proteins; D011994:Recombinant Proteins; D051545:Cyclooxygenase 1; D051546:Cyclooxygenase 2