PURIFICATION AND PARTIAL CHARACTERIZATION OF GLYCERALDEHYDE-PHOSPHATEDEHYDROGENASE FROM ELECTRIC ORGAN OF ELECTROPHORUS-ELECTRICUS (L.)

The glyceraldehyde-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) was pu rified to homogeneity from electric organ of Electrophorus electricus (L.) by a hydrophobic chromatography method on deacetylcolchicine-Seph arose. The purification resulted in a 162 fold increase in specific ac tivity of the GAPDH and final yield was approximately 37%. The purifie d enzyme showed a single band in SDS-PAGE, with an apparent molecular mass of 36 kDa. The purity of the colchicine-Sepharose isolated materi al was analysed by isoelectrophocusing and immunoblotting using a hete rologous rabbit serum anti-GAPDH. Sequence analysis of the 40-N-termin al amino acids, determined by Edman degradation, revealed its identity to other GAPDHs proteins being the largest number of identical amino acids to lobster (92.5%), rabbit muscle (85%) and human liver (80%) GA PDH.