Purification and characterization of human muscle pyruvate kinase

R. N. Harkins, J. A. Black, M. B. Rittenberg

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23 Scopus citations


The M1 isozyme of pyruvate kinase has been purified from human psoas muscle in a seven-step procedure. Fractionally by ammonium sulfate precipitation, heat treatment, acetone precipitation, diethylaminoethyl cellulose batchwise treatment followed by chromatography on carboxymethyl cellulose and Sephadex G-200 gave a product with a specific activity of 383 U/mg representing a 294-fold purification with a yield of 11%. The product formed orthorhombic crystals and was homogeneous on polyacrylamide gel electrophoresis with and without sodium dodecyl sulfate, sedimentation velocity, sedimentation equilibrium, and immunodiffusion. The purified enzyme has a molecular weight of 240700 and has a sedimentation coefficient (s20,w) of 10.04 S. It contains four subunits with identical molecular weights of 61 000. No free N-terminal amino acids could be detected. Antibody prepared against the purified human M1 isozyme does not cross-react by immunodiffusion or enzyme inactivation with the human erythrocyte isozyme and in the reverse experiment antibody prepared against human erythrocyte pyruvate kinase does not cross- react with the purified M1 isozyme. The amino acid composition of the M1 isozyme is presented.

Original languageEnglish (US)
Pages (from-to)301-307
Number of pages7
JournalCanadian journal of biochemistry
Issue number4
StatePublished - 1977

ASJC Scopus subject areas

  • Medicine(all)


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