Glyceraldehyde-3-phosphate dehydrogenase. Investigation on the regions responsible for self-assembly of subunits

Comp Biochem Physiol B. 1987;87(2):391-401. doi: 10.1016/0305-0491(87)90158-1.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / analysis
  • Animals
  • Carps / metabolism*
  • Chromatography, Gel
  • Chromatography, Paper
  • Cyanogen Bromide
  • Cyprinidae / metabolism*
  • Glyceraldehyde-3-Phosphate Dehydrogenases / analysis*
  • Iodobenzoates
  • Macromolecular Substances
  • Muscles / enzymology*
  • Peptide Mapping
  • Protein Conformation
  • Sulfhydryl Reagents
  • Trypsin

Substances

  • Amino Acids
  • Iodobenzoates
  • Macromolecular Substances
  • Sulfhydryl Reagents
  • 2-iodosobenzoic acid
  • Glyceraldehyde-3-Phosphate Dehydrogenases
  • Trypsin
  • Cyanogen Bromide