Transport of catecholamines by resealed chromaffin-granule ‘ghosts’

Abstract

A method is described for the preparation of resealed chromaffin-granule ‘ghosts’. The lysis and rapid purification procedures provide ‘ghosts’ in approximately 70% yield from crude granules; the preparation contains 0.1μmol of catecholamine/mg of protein (as compared with 2.8μmol/mg in unlysed granules), of which about one third is inside the ‘ghosts’. The ‘ghosts’ retain their ability to accumulate catecholamines, a process dependent on Mg-ATP and inhibited by reserpine, and a simple assay for this transport is described.