Glyceraldehyde-3-phosphate dehydrogenase. Investigation on the regions responsible for self-assembly of subunits.
1. In glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (EC 1.2.1.12) the four S-loop form the core of the tetramer. 2. Amino acid sequence of the S-loop of the regions of GAPDH from carp muscle was established through the analysis of tryptic digests of the enzyme treated alternatively with bromocyanate and o-iodosobenzoic acid. 3. Enzyme had been oxidized with performic acid. After treatment with trypsin the peptide mixture was fractionated into fragments. 4. CNBr cleavage of this enzyme was performed after S-carboxymethylation. The respective cyanogen bromide fragments have been isolated and characterized. 5. The procedure of protein fragmentation by o-iodosobenzoic acid used to split tryptophanyl peptide bonds. 6. Each peptide obtained after enzymatic or chemical fragmentation was purified to homogeneity by Bio-Gel or Sephadex chromatography, high voltage electrophoresis and descending paper chromatography and characterized by electrochromatography, N- and C-terminal sequence and amino acid composition. 7. The results are compared with those obtained from studies on GAPDH from other sources.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002347:Carps / Q000378:metabolism*; D002850:Chromatography, Gel; D002854:Chromatography, Paper; D003488:Cyanogen Bromide; D003530:Cyprinidae / Q000378:metabolism*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000032:analysis*; D007463:Iodobenzoates; D046911:Macromolecular Substances; D009132:Muscles / Q000201:enzymology*; D010449:Peptide Mapping; D011487:Protein Conformation; D013439:Sulfhydryl Reagents; D014357:Trypsin
D000596:Amino Acids; D007463:Iodobenzoates; D046911:Macromolecular Substances; D013439:Sulfhydryl Reagents; C008154:2-iodosobenzoic acid; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D014357:Trypsin; D003488:Cyanogen Bromide