PURIFICATION AND IMMUNOLOGICAL CHARACTERIZATION OF ACID BETA-GALACTOSIDASE FROM DOG LIVER

1. Dog liver acid beta-galactosidase was isolated in high yield and pu rified to homogeneity using a series of chromatographies on Con A Seph arose, decyl-agarose, anion-exchange HPLC and gel-filtration HPLC. 2. Non-denaturing gel filtration by HPLC gave a single homogeneous peak c orresponding to molecular mass of 180-190 kDa. During SDS PAGE analysi s, the single peak dissociated into a major band corresponding to mole cular mass of 32 kDa with minor bands at 18 and 13 kDa. 3. Polyclonal antibodies raised against the purified enzyme immunoprecipitated beta- galactosidase activity specifically from dog liver extracts and recogn ized a single 32 kDa band in Western blot analysis of dog tissue homog enates. This antibody did not crossreact with any protein band in tiss ue homogenates from other species examined except cat. 4. Western blot analysis of tissue extracts from dogs affected with GM1-gangliosidosi s showed the presence of a 32 kDa band similar to that of controls.