Ciliary membrane tubulin and associated proteins: a complex stable to Triton X-114 dissociation.
When either membranes from scallop gill cilia or reconstituted membranes from the same source are solubilized with Triton X-114 and the detergent is condensed by warming, no significant fraction of any major membrane protein partitions into the micellar detergent. Rather, most of the membrane lipids condense with the detergent phase, forming mixed micelles from which nearly pure lipid vesicles may be produced by adsorption of detergent with polystyrene beads. One minor membrane protein, with a molecular weight of about 20 000, is associated consistently with these vesicles. The aqueous phase contains a fairly homogeneous protein-Triton X-114 micelle sedimenting at 2.6 S in the analytical ultracentrifuge. Sucrose gradient velocity analysis in a detergent-free gradient indicates moderate size polydispersity but constant polypeptide composition throughout the sedimenting protein zone. Sucrose gradient equilibrium analysis (also in a detergent-free gradient) results in a protein-detergent complex banding at a density of 1.245 g/cm3. Sedimentation of the protein-detergent complex in the ultracentrifuge, followed by fixation and normal processing for electron microscopy, reveals a fine, reticular material consisting of 5-10-nm granules. These data are consistent with previous evidence that membrane tubulin and most other membrane proteins exist together as a discrete lipid-protein complex in molluscan gill ciliary membranes.
D000818:Animals; D002923:Cilia / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology*; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D005880:Gills; D046911:Macromolecular Substances; D008565:Membrane Proteins / Q000378:metabolism*; D008970:Molecular Weight; D008974:Mollusca; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology*; D013501:Surface-Active Agents / Q000494:pharmacology*; D014404:Tubulin / Q000378:metabolism*
D003902:Detergents; D046911:Macromolecular Substances; D008565:Membrane Proteins; D013501:Surface-Active Agents; D014404:Tubulin; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40