The specific association of a phosphofructokinase isoform with myocardial calcium-independent phospholipase A2. Implications for the coordinated regulation of phospholipolysis and glycolysis

S. L. Hazen, R. W. Gross

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Abstract

We have demonstrated previously that myocardial cytosolic calcium- independent phospholipase A2 is a 40-kDa polypeptide regulated by ligand- modulated protein-protein interactions (Hazen, S. L., and Gross, R. W. (1991) J. Biol. Chem. 266, 14526-14534). We now demonstrate that an 85-kDa polypeptide which possesses sequence homology to and chemical, physical, immunological, and chromatographic similarities with phosphofructokinase (PFK) specifically interacts with the 40-kDa phospholipase A2 catalytic subunit and represents the putative protein regulatory element identified in previous work. Multiple independent lines of evidence document the association between the 85-kDa phosphofructokinase isoform and the 40-kDa myocardial cytosolic calcium-independent phospholipase A2 catalytic polypeptide, including 1) the coelution of the 85- and 40-kDa polypeptides which migrate as a 400-kDa complex during gel filtration chromatography, 2) the stoichiometry between the 85- and 40-kDa polypeptides which corresponds to a complex comprised of a tetrameric PFK isoform and a 40-kDa phospholipase A2 catalytic polypeptide, 3) the demonstration that the 85-kDa phosphofructokinase isoform acts as a specific and reversible affinity adsorbant for myocardial cytosolic phospholipase A2 catalytic activity, 4) the immunoprecipitation of myocardial cytosolic phospholipase A2 activity utilizing chicken anti-rabbit skeletal muscle PFK IgG, 5) the specific release of phospholipase A2 from ATP-agarose after formation of a ternary complex comprised of allosteric modifiers of phosphofructokinase, and 6) the selective attenuation of the denaturation of purified homogeneous calcium- independent cytosolic phospholipase A2 with PFK. Collectively, these results demonstrate the highly specific association of a phosphofructokinase isoform with myocardial cytosolic calcium-independent phospholipase A2 and suggest a novel biochemical mechanism underlying the coordinated regulation of phospholipolysis and glycolysis previously observed in myocardium and in other mammalian tissues.

Original languageEnglish
Pages (from-to)9892-9900
Number of pages9
JournalJournal of Biological Chemistry
Volume268
Issue number13
StatePublished - Jan 1 1993

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