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Rapid purification of pig heart fumarase by general ligand chromatography

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Abstract

A rapid method for the purification of fumarase from pig heart muscle has been developed using general ligand chromatography with adenosine triphosphate as ligand. Fumarase exhibited distinctive elution patterns from several types of nucleotide-agarose matrices, which may prove of value in distinguishing putative isozymic forms. Fumarase purified from both soluble and particulate fractions of cardiac tissue appeared to be identical in terms of subunit molecular weight, electrophoretic mobility on cellulose acetate, substrate kinetics, and inactivation by several inhibitors. When fresh cardiac tissue was suspended in sucrose medium, both fumarase and citrate synthase were released from the mitochondria to about the same extent (10%). However, fumarase release was increased approximately three-fold, while the release of titrate synthase increased only slightly, when tissue which had been frozen and thawed was suspended in sucrose medium.

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Authors and Affiliations

  1. Pre-Clinical Science Unit, Veterans Administration Medical Center, 4500 South Lancaster Road, 75216, Dallas, Texas

    Laura A. Halper & Paul A. Srere

  2. University of Texas Health Science Center at Dallas, 5323Harry Hines Boulevard, 75235, Dallas, Texas

    Laura A. Halper & Paul A. Srere

Authors
  1. Laura A. Halper
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  2. Paul A. Srere
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Halper, L.A., Srere, P.A. Rapid purification of pig heart fumarase by general ligand chromatography. Journal of Solid-Phase Biochemistry 4, 1–13 (1979). https://doi.org/10.1007/BF02991803

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