CONVERSION OF β-N-ACETYLHEXOSAMINIDASE B TO A BY GOLGI COMPLEX OF RAT PAROTID GLAND

Publisher Summary This chapter presents an experiment that was conducted to convert β-N-acetylhexosaminidase B to A by the Golgi complex of the rat parotid gland. In this experiment, the homogenate was treated with cetyltrimethyl ammonium bromide at a final concentration of 0.4% at 4°C for 60 min. The mixture was centrifuged at 25,000 g for 40 min, and the supernate was concentrated with 80% saturation of ammonium sulfate. The sediment was dialyzed and applied to CM-cellulose column chromatography, which was equilibrated with 0.01 M phosphate buffer, pH 7.4. Hexosaminidase B was eluted with the buffer containing 0.125 M sodium chloride. In the isolating procedure of the Golgi apparatus, the supernate at 100,000 g for 30 min was boiled for 10 min. The boiled material was centrifuged at 15,000 g for 30 min, and the supernate was used as boiled juice.