Ronald K. Scheule, Betty J. Gaffney
Oct 1, 1981
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Influential Citations
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Journal
Analytical biochemistry
Abstract
Abstract Triton X-100 is a neutral detergent which is often employed for reconstitution of lipid-protein membranes. It is, however, difficult to remove entirely from the final reconstituted membrane preparation. We have analyzed the molecular components of the Triton X-100 complex of molecules that remain in reconstituted membranes which are prepared by, first, Triton X-100 solubilization and, second, by detergent removal with polystyrene beads and gel filtration. The analysis was performed with radioactive Triton X-100 and by silica gel chromatography. It showed that the components of Triton X-100 that are difficult to remove from a membrane are the high- and low-molecular-weight ones. Using only an intermediate-molecular-weight, radioactive Triton, lipids and Sindbis virus envelope proteins have been solubilized and used to prepare reconstituted membranes which retain half as many Triton/lipid molecules, or one-fifth as many Triton/protein molecules as do vesicles prepared with complete Triton X-100. The membranes prepared with fractionated Triton are also of more homogeneous size, by gel chromatography, than are membranes prepared with unfractionated Triton.