| Abstract: | Affinity chromatography, a separation technique which makes use of biospecific properties, is well established for the separation of molecules in solution. We applied this method to the subfractionation of biomembranes. Using a microsomal fraction mainly consisting of plasma membrane from rabbit or calf thymocytes, 20–40% of the protein adhered specifically to concanavalin A-Sepharose, whereas the majority of the membrane vesicles were recovered from the effluent. The adherence involved the binding of Con A to membranes, as addition of the hapten sugar α-methyl-mannoside completely prevented separation. The fractions which bound to Con A-Sepharose could be eluted by combining mechanical forces with the addition of α-methyl-mannoside. All fractions exhibited the same vesicular appearance and were identical with respect to the phospholipid cholesterol ratio. The method proved to be highly reproducible and it offers a possible way for the subfractionation of membranes according to their biospecific structure. |
