A new radioactive diazofluorene based heterobifunctional reagent.

Synthesis of a radioactive photoactivable heterobifunctional reagent, Noxysuccinimide ester of 2-[14C]glycyl carboxy-9-diazofluorene is described. This reagent on photolysis gives rise to a reactive carbene which rapidly inserts into solvents like methanol. The probe can be easily linked to aldolase which on photolysis gives rise to aldolase dimer, trimer and tetramer depending on the density of linked probe. This probe has also been linked to concanavalin A. The radioactive concanavalin A so obtained was incubated with erythrocyte ghosts and photolysed. The membrane protein analysis by gel electrophoresis indicated that concanavalin A has been covalently crosslinked to band 3.

Cholesteryl-(2'-hydroxy)-ethyl ether—A potential cholesterol substitute for studies in membranes.

The yeast sterol auxotroph GL-7, which grows well on ergosterol and cholesterol, was used to study the ability of cholesteryl-(2'–hydroxy)-ethyl ether to substitute for cholesterol. In this compound the 3j3-hydroxyl group of cholesterol is replaced by ethylene glycol and the resulting ether still retains the amphiphilic character of cholesterol. Cholesteryl-(2Lhydroxy)-ethyl ether was found to support the growth of GL-7 as effectively as cholesterol. Crystal violet permeability and membrane order parameter determined using a spin label were similar for cells grown on these sterols. The ability of such ethylene glycol derivatives to substitute for cholesterol in both artificial and natural membranes should help in designing suitable spacers through which molecules can be linked to cholesterol without affecting the normal function of cholesterol in membranes. This in turn should prove useful in studies with surface -modified liposomes.