Isolation and sequence determination of an active site peptide of rabbit muscle pyruvate kinase

Guillermo Bezares, Jaime Eyzaguirre, Maria Victoria Hinrichs, Robert L. Heinrikson, Ilene Reardon, Robert G. Kemp, Steven P. Latshaw, Sergio Bazaes

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)


Rabbit muscle pyruvate kinase was inactivated by 2′,3′-dialdehyde ADP with the incorporation of one molecule of reagent per enzyme subunit. The inactivated protein was digested with trypsin after reduction and carboxymethylation. The labeled peptide was isolated by gel filtration and further purified by HPLC. The peptide was sequenced both by liquid-phase and gas-phase automatic Edman degradation. A 34-residue peptide was obtained. This peptide is identical to a tryptic peptide labeled with trinitrobenze-nesulfonate, isolated and sequenced by Johnson et al. (Biochem. Biophys. Res. Commun. (1979) 90, 525-530) from bovine muscle pyruvate kinase. Available evidence suggests that dialdehyde ADP labels the enzyme at the same lysine in position 25 of the peptide, as found by Johnson et al. The high homology between the isolated peptide and regions of other pyruvate kinases from low to high eukaryotes supports the idea that this peptide is related to the enzyme active site.

Original languageEnglish
Pages (from-to)133-137
Number of pages5
JournalArchives of Biochemistry and Biophysics
Issue number1
Publication statusPublished - 15 Feb 1987

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology


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