Purification and partial characterization of glyceraldehyde-phosphate dehydrogenase from electric organ of Electrophorus electricus (L.).
The glyceraldehyde-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) was purified to homogeneity from electric organ of Electrophorus electricus (L.) by a hydrophobic chromatography method on deacetylcolchicine-Sepharose. The purification resulted in a 162 fold increase in specific activity of the GAPDH and final yield was approximately 37%. The purified enzyme showed a single band in SDS-PAGE, with an apparent molecular mass of 36 kDa. The purity of the colchicine-Sepharose isolated material was analysed by isoelectrophocusing and immunoblotting using a heterologous rabbit serum anti-GAPDH. Sequence analysis of the 40-N-terminal 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%) GAPDH.
Giovanni-De-Simone
S
S
Departamento de Biologia Celular e Molecular, Universidade Federal Fluminense, Niterói, RJ, Brasil. dsimone@gene.dbbm.fiocruz.br
Hassón-Voloch
Batista-e-Silva
C
C
Nery-da-Matta
A
A
D000595:Amino Acid Sequence; D000818:Animals; D002846:Chromatography, Affinity; D004331:Drosophila melanogaster / Q000201:enzymology; D004557:Electric Organ / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D004593:Electrophorus; D004926:Escherichia coli / Q000201:enzymology; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D008121:Nephropidae; D010446:Peptide Fragments / Q000737:chemistry; D011817:Rabbits; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013824:Thermus / Q000201:enzymology; D014246:Trichomonas vaginalis
D010446:Peptide Fragments; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases