Imaging of reconstituted purple membranes by atomic force microscopy.
The organization of bacteriorhodopsin (bR) within reconstituted purple membranes (RPM) was examined using atomic force microscopy (AFM). Five reconstituted species were examined: RPM 3 (bR/native polar lipids/dimyristoylphosphatidylcholine (DMPC) in a 1:9:14 molar ratio), RPM 4 (bR/native polar lipids in a 1:7 molar ratio), RPM 5 (bR/native polar lipids/1,2-di-O-phytanyl-sn-glycerol in a 1:3.5:6.1 molar ratio), RPM 6 (bR/native polar lipids/1,2-di-O-phytanyl-sn-glycero-3-phosphocholine in a 1:3.5:4.9 molar ratio), and RPM 7 (bR/native polar lipids/1,2-diphytanoyl-sn-glycero-3-[phospho-L-serine] in a 1:3.5:4.6 molar ratio). RPM 3 patches adsorbed onto mica exhibit domains of crystallized bR trimers arranged in a hexagonal packing structure, similar to those found in native purple membrane (NPM). These domains are enclosed by DMPC-rich regions. RPM 4 patches were observed to have larger domains of crystallized bR, with trimer orientation 30 degrees different from that found in NPM. The bR-rich domains are enclosed by a large, protein-free, lipid-rich region. The topography of RPM 5 was difficult to resolve as the surface had no discernable patterns or structure. The topographies of RPM 6 and 7 were similar to that found in RPM 3 in that higher domains were formed within the patch adsorbed onto mica. They may contain protein-rich regions, but clear images of protein arrangement could not be obtained using AFM. This may be a result of imaging limitations or of the lack of organization of bR within these domains.
Kim
David T
DT
Department of Chemical Engineering, University of California, 201 Gilman Hall, Berkeley, CA 94720-1462, USA.
Blanch
Harvey W
HW
Radke
Clayton J
CJ
D000327:Adsorption; D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000648:ultrastructure; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D008055:Lipids / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry*; D018625:Microscopy, Atomic Force / Q000379:methods; D010715:Phosphatidylglycerols / Q000737:chemistry; D011506:Proteins / Q000737:chemistry; D018717:Purple Membrane / Q000737:chemistry* / Q000145:classification / Q000648:ultrastructure
D008055:Lipids; D008081:Liposomes; D010715:Phosphatidylglycerols; D011506:Proteins; D001436:Bacteriorhodopsins; D004134:Dimyristoylphosphatidylcholine